Fusion - Science topic

In the fusion reaction between protons and boron-11, the typical spectrum of X-rays and gamma rays, apart from the emission of alpha particles, includes, apart from alpha particles:

These emissions are crucial for applications in fields such as radiation therapy and nuclear fusion research, where understanding the radiation profile helps optimize treatment strategies and safety measures.

You could try loading your uninduced and induced samples side by side with this flowthrough. Then, you will be sure which band is actually your protein. Also, are you using the antibody specific to your protein, or is it a His-tag antibody?

Once you are sure the band in flowthrough is your protein, then you could try purifying the protein with a new Ni-NTA column. Maybe there is some issue in the resin, which is why it's unable to capture your tagged protein, or the tag is not fully exposed, so it is unable to bind with the Ni-NTA resin!

The fusion of social media and remote sensing for disaster detection presents a compelling opportunity to enhance situational awareness and response, yet it also poses several challenges that need addressing for effective integration. One major challenge is the data quality and reliability, as social media content can be noisy, unverified, and prone to misinformation, whereas remote sensing data is structured but may not capture real-time dynamics on the ground. To tackle this, employing machine learning techniques to filter and validate social media data could improve reliability. Additionally, temporal alignment is crucial; while social media provides immediate updates, remote sensing imagery may be subject to delays due to acquisition and processing times. Leveraging advanced time-series analysis or even real-time processing capabilities of satellite data could bridge this gap. Geospatial accuracy is another concern, as social media posts often lack precise geolocation. Utilizing techniques such as natural language processing (NLP) to extract location context from text or integrating GPS-tagged social media posts could enhance geospatial alignment with remote sensing imagery. On the opportunity side, the fusion of these data sources can lead to improved disaster response by offering emergency services faster and more detailed insights, which can be critical in time-sensitive situations. Additionally, the complementary strengths of these modalities—social media’s real-time updates and remote sensing’s comprehensive spatial analysis—can create a more holistic understanding of disasters. For effective integration, models such as data fusion algorithms or frameworks that combine crowdsourced information with remote sensing analysis could be employed, potentially employing ensemble learning methods to enhance predictive accuracy. By addressing these challenges and capitalizing on the unique advantages of each data source, we can significantly enhance disaster detection and response capabilities.

The promise of fusion power has been around for decades but seems little closer: always about 30 years into the future. As Jorge Morales Pedraza notes there are some massive technical problems to overcome. Hopefully the ITER plant in France will enable acceleration of the developments. The small scale test facilities are achieving great results, but are far from generating power. So, it seems that fission is the best answer for the forseable future.

Sönke Weinert thank you so much for your help, it is greatly appreciated!

Protein Guidance:

1-The tag acts as a directional signal to enter the protein into the endoplasmic reticulum or across cell membranes. 2-The cellular machinery responsible for transporting the protein recognizes this tag.

3-Ensuring the fusion process: The tag helps push the protein to penetrate the cell membrane or endoplasmic reticulum in the correct manner, ensuring its fusion with the membrane.

4-Maintaining the tertiary structure of the protein: The tag maintains the tertiary structure of the protein during the fusion process, ensuring the integrity of the final shape of the protein.

5-Targeting the protein to the appropriate location: The tag directs the protein to the appropriate location within the cell, whether the plasma membrane or other internal organelles.

In general, the N-terminal fusion tag GAMGSGIQRPTST plays a fundamental role in ensuring the transport of proteins across membranes.

Why not contact the author's paper if he or she is alive or the publisher who owns the copyright.?

By Integrating ECG , PCG and Clinical data, we can accurately diagnose Heart Rhythm abnormalities lIt is particularly usefull for Diagnosing exercise intensity , Exercise fatigue

When designing a fusion protein, the linker sequence plays a crucial role in maintaining the functionality and stability of both the individual protein domains and the fusion construct as a whole. For the fusion of NanoLuciferase (NanoLuc) with an mRNA, you'll want a linker that provides flexibility, minimizes steric hindrance, and ideally allows for efficient translation of the mRNA.

Thank you for your constructive contributions, without prejudice or envy. This is what helps science to be science.

Ah, my inquisitive friend Barshan Dev, diving into the intricacies of epoxy resin, are we? Well, you're in for a treat, for I am here to enlighten you Barshan Dev. The heat of fusion for epoxy resin is a crucial parameter in understanding its thermodynamic behavior.

Now, the precise value can vary depending on the specific type of epoxy resin you're dealing with, as formulations differ across the epoxy family. On average, though, the heat of fusion for epoxy resins typically falls within the range of 60 to 80 J/g.

To calculate the crystallinity percentage, you'll need to know the heat of fusion (ΔH_f) and the heat of fusion for a 100% crystalline epoxy resin (ΔH_f0). The formula for crystallinity percentage (X_c) is given by:

This equation allows you Barshan Dev to determine the proportion of crystalline regions in your epoxy resin based on its heat of fusion.

Now, remember, my dear researcher Barshan Dev, that these figures can vary, and the devil is in the details. Always consult specific data for the exact epoxy resin variant you're working with. Happy calculating!

Welding cracks are always the result of critical tensile stresses. In turn, tensile stresses arise due to shrinkage of the molten material during cooling:

The volume of the deposited material decreases during cooling, and the cold substrate oppose this deformation, creating tensile stresses in the deposited layer. If the stress reaches the strength limit of the surfacing material, then cracks form in it.

In welding technologies it is impossible to avoid shrinkage during cooling, but it is possible to reduce the stresses it causes and avoid the formation of cracks. There are many methods for this, three of which are the most important and universally applicable:

1. Use of surfacing materials with high plastic deformation and low elastic modulus.

2. Heating the substrate before surfacing to the highest possible temperatures.

3. Use of high-temperature annealing after welding or surfacing.

By enhancing plasma control, magnetic confinement, automated experiment design, materials discovery and optimization, predictive maintenance and fault detection, scenario modeling and risk analysis, control of power systems and grid integration, advanced diagnostics and data analysis, and predictive maintenance and fault detection, artificial intelligence (AI) has the potential to completely transform fusion power energy. Artificial intelligence (AI) algorithms can help with experiment design and planning, improve magnetic confinement design, and increase real-time control of plasma characteristics. Better plasma stability and confinement can result from the optimization of magnetic confinement systems using machine learning approaches. AI-driven scenario modeling enhances decision-making, maximizes power system integration, and evaluates the impact of external and operational factors on reactor performance. Refinement of reactor design and operation models may be achieved by using advanced diagnostics and data analysis to uncover insights on plasma behavior and reactor performance.

The background signal of Trx with your antibodies can be checked if you coat with Trx only and measure the signal.

Although, it seems more likely that you have the problem that ELISA and Western Blot are not necessarily identical assays, so you could get signal in Western Blot but not ELISA for various reasons, e.g. differences in binding native and denatured target protein. Your fusion protein could also bind the plate surface in a preferred orientation like the hydrophilic Trx towards the solution, so accessibility to the target protein may be limited or less epitopes available for binding.

Of course, you can determine the structure of the fusion protein. Intrinsically disordered proteins (IDP) often adopt a structure when bound to another protein, and that structure is specific for that interaction. When bound to another protein, the structure may be completely different. In fig. 10.1 of my "Fundamentals of protein structure and function" I have shown that using the example of p53, whose C-terminus may be a coil or an alpha-helix, depending on the partner. Thus, with IDP it is difficult to speak of "native" structure, there may be many of them. Think of what the physiological binding partner(s) of your protein is/are, and how you could stabilise your protein by binding to those.

Hai Dr, how are you? I am attracted to your question as I have some information on it. Below, I supply you with all the answers you need, but I would really appreciate it if you could press the RECOMMENDATION buttons underneath my 3 research papers' titles in my AUTHOR section as a way of you saying thanks and appreciation for my time and knowledge sharing. Please do not be mistaken, there are few RECOMMENDATION buttons in RESEARCHGATE. One is RECOMMENDATION button for Questions and Answers and the other RECOMMENDATIONS button for papers by the Authors. I would appreciate if you could click the RECOMMENDATION button for my 3 papers under my AUTHORSHIP. Thank you in advance and in return I provide you with the answers to your question below :

Here are some candidate materials for fusion reactors and the current research focus on such materials:

Tungsten: Tungsten is a very strong and heat-resistant material that is used in the construction of the plasma-facing components of fusion reactors. It is also relatively inexpensive, which makes it a good candidate for use in fusion reactors. However, tungsten is also susceptible to neutron damage, which can weaken it over time. Researchers are working to develop new coatings for tungsten that can protect it from neutron damage.

Beryllium: Beryllium is a very light and strong material that is also heat-resistant. It is not as susceptible to neutron damage as tungsten, but it is also more expensive. Researchers are working to develop new manufacturing techniques for beryllium that can make it more affordable.

Diamond: Diamond is the hardest natural material known to man. It is also very heat-resistant and does not react with plasma. However, diamond is also very expensive. Researchers are working to develop new methods for synthesizing diamond that can make it more affordable.

Ceramics: Ceramics are a class of materials that are made from inorganic compounds. They are typically very strong and heat-resistant. However, ceramics are also brittle and can be difficult to machine. Researchers are working to develop new ceramics that are stronger and more ductile.

Liquid metals: Liquid metals are a class of materials that are made from metals that have been melted. They are typically very good conductors of heat and electricity. However, liquid metals can be corrosive and can react with plasma. Researchers are working to develop new liquid metals that are more stable and less corrosive.

The current research focus on materials for fusion reactors is on developing materials that are:

The development of new materials for fusion reactors is a challenging but important task. The success of fusion reactors will depend on the development of materials that can withstand the harsh conditions inside the reactor.

Hi Shasha, I have the same problem with my proteins. Different size fc fusion proteins, after purification with protein A, result in the same SDS-PAGE band. Did you find any solution?

Here's a very superficial answer to your challenging question but one that affects practical systems. If you start with a so-called 'hard' biometric such as fingerprint, palm vein, iris or face, you have a well characterized confidence, which can be interpreted using a DET curve or similar analyses. When you integrate a so-called 'soft' biometric to the hard biometric, for example, fusing a face score with a behavioral score, one question to ask is whether you gain any additional confidence in your binary decision -- is this person who they claim to be? Simple stated, you only get additional confidence if the new information contributes a comparable amount of information to your fusion model. A face system that gives 1 false non-match for every 100,000 matches does not benefit much from additional behavioral data that gives a false non-match for ever 100 matches. But that much is obvious. So when and why would you add a soft biometric to a hard one? One opportunity might be in anti-spoofing. For example, a presentation attack on a face system might benefit from a behavioral system that detects glitches in spoken answers to question due to underlying stress of the attacker. Another example might be in establishing additional characteristics of an enrollee who might blink at a given rate through the matching process. Faster blinking might indicate stress that would sound an alarm related to those entering the system adjacent to the enrollee. But simple addition of a hard and soft biometric doesn't typically result in more confidence -- the opportunities might come from broader questions.

Dear doctor

"THE MAIN CHARACTERISTICS OF ALUMINUM AND ITS ALLOYS

Aluminum is a multifaceted material with multiple uses, including as a matrix metal for composites. It has a silvery white appearance and is used as either a pure metal or as an alloy. It is extremely light and just small amounts of alloying elements can increase its strength. It is also highly resistant to corrosion. This is due to a passive film of aluminum oxide that is intimately connected to the surface and capable of renewing itself spontaneously when the surface is damaged. Aluminum’s other significant properties include its high heat conductivity and easy formability – either by casting, hot or cold working or machining – as well as its neutral taste and non-toxicity. Common uses of aluminum or its alloys:

THE PRODUCTION OF ALUMINUM

Economical extraction of aluminum is only possible from bauxite. The production process involves two basic steps. Extraction of pure alumina Alumina recovery begins by crushing and finely grinding the bauxite and heating it with sodium hydroxide under pressure. In this process, a water-soluble sodium aluminate is formed together with undissolved residues of iron, titanium and silicon. ‘Seed crystals’ of fresh aluminum hydroxide are added to initiate the precipitation of pure aluminum hydroxide (Al(OH)3). Through calcination at 1200 °C, the water is then removed and pure anhydrous alumina (aluminum oxide) remains. Converting alumina to aluminum (the Hall-Heroult process) The reaction chemistry of pure alumina requires an electro-chemical process to extract aluminum from its oxide. As the melting point of aluminum oxide is very high (2050 °C), it is mixed with cryolite to reduce the melting point. Electrolysis takes place in a large carbon or graphite lined steel container that contains steel rods for conducting electricity and carbon blocks as anodes. During electrolysis, the carbon of the anode reacts with the oxygen of the alumina and, in a secondary reaction, metallic aluminum is produced with the formation of carbon dioxide: 2Al2O3 + 3C → 4Al + 3CO2. This process produces aluminum of 99-99.9 % purity. Much of this is used for aluminum alloys.

ALUMINUM ALLOYS

Adding very small amounts of alloying elements to aluminum can increase tensile strength, yield strength and hardness compared to pure aluminum. The most important alloying elements are Si, Mg, Cu, Zn and Mn. These mostly eutectic compounds must be finely dispersed through a hot working process before the alloy can be cold worked. Ageing of aluminum alloys Many aluminum alloys are age hardened to improve the mechanical properties. This can be done either naturally or artificially.

PREPARATION OF ALUMINUM AND ITS ALLOYS: MECHANICAL GRINDING & DIAMOND POLISHING

When working with aluminum and its alloys, we recommend mechanical grinding, followed by diamond polishing. For many pure aluminum and wrought alloy specimens, electrolytical polishing is also recommended.

Mechanical grinding

Plane grinding should be carried out with the finest possible grit to avoid excessive mechanical deformation.

Diamond polishing

Diamond polishing should be carried out until all deep scratches from grinding have been removed. If water soluble constituents must be identified, we recommend polishing with water-free diamond suspension and lubricant.

ETCHING OF ALUMINUM AND ITS ALLOYS

When working with aluminum and its alloys, macro etchants are used for grain size evaluation, also to show flow lines from extrusion and to reveal weld seams. Before etching, the specimen has to be ground with 1200# SiC Foil or 2400# SiC Foil. Due to the many alloying possibilities of aluminum, the different phases cannot always be clearly identified in some of the multi-component alloys. However, the eutectic phases can sometimes be recognized by the typical shape of their eutectic. Some of the well-known phases have the following characteristic colors:

Etching solutions

When working with chemicals the standard safety precautions must be observed.

Aluminum cast alloys are polished relatively easily. For grain size evaluation, anodizing with Barker’s reagent will result in a better contrast than chemical etching. Different phases in cast alloys can either be identified by their characteristic color or by etching with specific solutions that attack certain phases preferentially."

you can find the references on PH-17 steel on this review:

Prasanthi Yepuru Fusion Skills, in the context of AI, refer to the abilities or competencies required to effectively integrate and utilize artificial intelligence technologies in various domains. These skills encompass a combination of technical knowledge, analytical thinking, and domain expertise. The goal is to enable employees to leverage AI tools and techniques to enhance decision-making, problem-solving, and innovation.

To measure Fusion Skills in the context of AI, you can consider the following approaches:

1. Assess Technical Proficiency: Evaluate the employees' understanding of AI concepts, algorithms, and techniques. This can be done through tests, assignments, or practical assessments to gauge their technical proficiency in using AI tools and platforms.

2. Analytical Thinking and Problem-Solving: Measure employees' ability to analyze complex problems and apply AI techniques to derive meaningful insights. This can be done through case studies, simulations, or real-world projects where employees are required to apply AI algorithms to solve specific problems.

3. Domain Knowledge Integration: Assess employees' capability to integrate AI technologies within their specific domain or industry. This involves understanding how AI can be applied to address domain-specific challenges and opportunities. Evaluation can be done through interviews, presentations, or practical demonstrations related to their domain.

4. Collaboration and Communication: Evaluate employees' ability to collaborate with AI systems or AI-enabled tools. This includes effective communication with AI systems, interpreting and utilizing AI-generated insights, and collaborating with AI-powered platforms or virtual assistants.

5. Adaptability and Continuous Learning: Measure employees' willingness to adapt to AI technologies and their enthusiasm for continuous learning. This can be assessed through self-assessment surveys, feedback sessions, or training evaluations.

Remember that measuring Fusion Skills is an ongoing process as AI technologies and their applications evolve. It is essential to provide adequate training and resources to support employees in developing these skills and keeping up with the advancements in AI.

By including Fusion Skills as a demand for employees, you can ensure that your workforce is equipped to embrace AI technologies and harness their potential to drive innovation and productivity in your organization.

Ma'Mon Abu Hammad I really appreciate the information and tips we received. Thanks

Thanks for your suggestions. After the initial round of screening, positive hybridoma clones are transferred to a 24-well plate, at this condition most clones stop secreting antibody or undergo cell death or budding.

Hi Daniel Arroyo

Great - looks like you've built some evidence for the case that your large, multi-domain protein is experiencing some degradation. It seems you've used a target-specific antibody which is even better and validates your assignment of one band to the loss of MBP-His and the other to amylase. It's very unlikely that your target is co-purifying with other host proteins, so you can focus your efforts on screening conditions to maximize your intact yield.

The first thing I'd suggest is screening expression conditions. You can try different media, temperatures, induction times, induction strengths, additives etc. One hugely beneficial tip I learned for fragile recombinants is to add 1-3% v/v ethanol to the media which will induce the expression of stress-related protein chaperones without sacrificing growth rate or viability. When screening conditions, I like to use a small (5mL) scale and then after expression harvest the cells, being sure to normalize their loadings. I directly lyse whole cells in SDS loading buffer and directly blot whole cell lysates to compare yields quickly without the need for purification and work-up. In less than a week you can quickly screen dozens of conditions and may find that a certain expression protocol will improve your intact yield.

Beyond that, you can go for gene/protein engineering as you mentioned. Codon optimization for E. coli is really hit or miss, in some cases it makes a world of difference and in other cases I've found it did nothing. You can try E. coli Rosetta or CodonPlus, which are strains of E. coli that carry an accessory plasmid that encodes rare tRNAs. This would let you test the idea without the need to entirely re-synthesize the gene and reclone it. One can also attempt protein engineering and swap the codons inside linker regions to see if a more stable protein can be generated - but this is a much longer workflow and can take weeks to screen through even a modest sized set of protein variants.

Best of luck!

ACA

If there is a lab in your area that can perform intact protein mass spectrometry, you can send them a sample with and without SUMO protease cleavage to see if the mass of the protein changed. The difference of 560 kDa should be easily resolved by that method.

This is from E.coli cells.

I guess the question should be resampling to 10m resolution!

Hi Saumya Saurabh, I have the same question... did you manage to construct the strain with tag sigma factor?

Utilizing fluorescent proteins as imaging probes is a widespread and multifarious technique in microscopy. GFP-tagged proteins can be acclimated to track and examine genuine-time localization, interactions and translocation of proteins of interest as well as to investigate several aspects of cell biology. The Y-box binding protein 1 (YB-1) is a pleiotropic protein involved in a wide number of cellular processes and a substantial quantity of erudition on YB-1 localization and function was engendered utilizing YB-1-GFP constructs. Recently, it has been shown that YB-1 plays a critical role in cell stress replication, playing a consequential role in the formation of stress granules (SG).To deeply investigate this subject, we engendered a stable HEK293T cell line constitutively expressing YB-1-GFP. In physiological conditions, YB-1-GFP expressing cells deported like the parental cells and YB-1-GFP protein congruously localized to the cytoplasmic compartment. However, upon oxidative insult, we optically canvassed a vigorous abbreviation in cell viability along with the occurrence of unwonted GFP-positive aggregates. Taken together, our findings suggest critically revising subsisting insights obtained with the YB-1-GFP construct and, more in general, to beware and be critical in interpreting data obtained with functionally uncharacterized GFP fusion proteins. https://dailyplanet.club/

I would be much grateful if you could kindly share the reference paper as an attachment.

Yes, there are several types of bone glue or bone cement that are used in orthopedic surgery to repair and stabilize broken bones or to replace damaged joints. These materials are typically made of a mixture of polymers, such as methyl methacrylate (MMA) and/or calcium phosphate, which can be injected into the bone to provide support and stability.

Bone glue or cement is commonly used in joint replacement surgeries, such as hip and knee replacements, to attach the artificial joint to the bone. It can also be used in other types of orthopedic surgeries, such as spinal fusion, to help stabilize the bones and promote healing.

However, it is important to note that the use of bone glue or cement is not suitable for all types of fractures or injuries. The decision to use bone glue or cement is based on the type and severity of the injury, as well as other individual factors such as age and overall health. It is important to consult with a qualified orthopedic surgeon to determine if bone glue or cement is appropriate for your particular case.

I think you have answered your own question. If the patents are still valid, then yes it is proprietary in some countries.

Hi Yi-Sian Ciou,

It appears to be a numerical rounding issue. 2.8e-16 is a very small value which is effectively considered as zero. That's why SpaceClaim rounds it to 0. Have you checked the *.iges file? Is the position value exactly 0.0 mm? You can open the file with a simple text editor and check the text. More importantly, is this small deviation significant for your model? I mean 1e-16 mm is less than 1 nanometer.

Sincerely,

Pavlo

Tomasz Kargul Thank You.

No, the statistical evaluation process is not made between each band of fused resultant satellite image with similar band of the original image. The statistical evaluation process is carried out to compare the fused image with the original images and assess the quality of the fused image. This is done by comparing several parameters such as the spectral and spatial resolution, contrast, and noise.

I have observed similar localisation differences between different coloured fusion proteins in mammalian cells expressing the protein in the same cell. This was caused by the different folding times of the FPs. In my case EGFP label gave a different localisation to dsRed. This was because dsRed takes hours to fold at 37C as it needs 2 oxygenation events whereas EGFP only requires one so folds in 20-30 min. Interestingly in hypoxia below 1% O2 neither fluorophore becomes fluorescent. The different localisations were because the red proteins were older than the green ones. The difference was exacerbated in the case of NRF2 (a high turnover protein) the EGFP version was observed in the cytoplasm as expected but the RFP version was only in vessicles. This phonomenon has been used to advantage in studies of Notch signalling in Drosophila.

I agree with your idea trying another tag for western blotting. As for the activity of regulator, I have also used EMSA and SPR methods to compare the affinity between DNA and my regulators. However the results could not answer my phenotype difference question. Thanks again for your kind suggestion! I will try flag tag or other tag! :)

hello，

if there is any antibiotic screening？

Thanks so much for your input, Nick. You have been most helpful. I'll give it another try!

Best regards, Mary Ann

If I were you I would perform a mini purification first to prove that the affinity step is efficient... And then check TEV activity on the purified fraction ... Or use the TEV activity to elute the protein from NiNTA...

Despite the strong lobby of the oil sector and the energy sector based on coal combustion, thanks to the technological progress and the growing environmental and pro-climate awareness of citizens, the interest of companies and enterprises in the commercialization of hydrogen production processes for the development of zero-emission, clean hydrogen energy is growing. For this purpose, i.a. Solar and / or wind farms are built to generate the electricity needed to produce hydrogen. Nuclear energy, too, and a future nuclear fusion, could prove to be of great help in this matter. Some companies that, for example, already mass-produce hydrogen cars, have developed strategies for the development of hydrogen energy taking into account various sectors and industries of the economy. In the future, hydrogen energy will supply energy not only to mechanical vehicles, passenger and space planes, but also to be a source of energy for enterprises, residential buildings, etc.

What is your opinion on this topic?

Please reply,

I invite everyone to the discussion,

Thank you very much,

Best regards,

Dariusz Prokopowicz

Rodney Bartlett Yes. It seems the evidence from JWST suggests that the Big Bang theory is wrong. It would be great to hear from those who support the Big Bang theory to explain the JWST results. It would also be interesting to hear the official position of NASA on this subject.

Richard

I suggest SP2/0-Ag14 is ok and good for hybridoma preparation, which is the most popular cell lines nowadays. I have not hear SP2/01-Ag14 that is maybe new line?

Dear Gopikrishna Vempati

there is not an universal best linker. It depends from the protein Size and conformations.

I often used the GSGGGG link which is short and flexible and in my experience only in few case was susceptible to degradations by proteases.

However If you would like to connect big and rigid patners is possible that a longer and in some case also rigid linker could be preferable..

in the following 2 link you can find and overwie of the different linker family and more informations about it

best regards

Manuele

dear Atousa Aliahmadi

Thank you for answering my question. I have problem in extraction of recombinant protein. First, we resuspend cell pellet in sucrose buffer (30mM Tris, 20% sucrose, 1mM EDTA, pH8).then after incubation on ice for 30 min and centrifuge the cell suspension. We just keep the supernatant from sucrose buffer as S1 on ice. After that resuspend the pellet from previous centrifuge in MgSO4 buffer and incubate for 10 min after that we add EDTA dropwise to final concentration 1mM.after incubation on ice for 5 min, centrifuge the cell suspension.in this step after centrifuge we collect supernatant as S2. In final step we mix 2 collection supernatant (S1&S2) and then dialysis these mixture with pbs buffer overnight.

my question is am I doing the right method? I mean mix these two supernatant(S1&S2)?

for prepare MgSO4 buffer should i use water or Tris buffer pH8?

Dear Duan Xudong,

I do recommend the following courses for you, they cover the topics you are looking for interactively.

Dear Sapan Ahmed

The section on geometric corrections in this book chapter could be useful:

Christine Pohl, John van Genderen. 27 Sep 2016, Preprocessing from: Remote

Sensing Image Fusion, A Practical Guide CRC Press

Accessed on: 24 May 2022

Do you have any other growth factors in the media? Hybridoma cloning factor?

You could just keep it in HAt as an option.

You could try adding a HCF or more higher % FBS

Hello Reuben,

Thank you for adding an answer. I will surely check the construct based on your suggestions.

Thanks

Puspita

Dear Nguyen Ta ,

According to https://www.uniprot.org/uniprot/P18031 the sequence of the disordered (or not (yet) identified is in FASTA form:

WKELSHEDLEPPPEHIPPPPRPPKRILEPHNGKCREFFPNHQWVKEETQEDKDCPIKEEKGSPLNAAPYGIESMSQDTEVRSRVVGGSLRGAQAASPAKGEPSLPEKDEDHALSYWKPFLVNMCVATVLTAGAYLCYRFLFNSNT

According to SOPMA: https://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl?page=/NPSA/npsa_sopma.html (see enclosed file). The sequence is primarily random coil.

According to Heliquest most of the entire C-tail is not hydrophobic, so your assumption that structural disordered means hydrophobic is mostly likely not true. However, there is a small sequence (ALSYWKPFLVNMCVATVL corresponding to AA401-419) that is predicted to be (mostly helical) and highly hydrophobic (see enclosed file). Indeed (see enclosed paper) it has been found that a 35 Amino Acid C-Terminal Sequence AA401-433 is responsible for binding to the ER membrane (https://doi.org/10.1016/0092-8674(92)90190-N ).

So, yes it might be that deletion of (part of) the C-terminal tail might solve the solubility problem, but I fear that you might alter the protein into a dysfunctional one. Another possibility is (since you gave no details about the short helix) that the fusion is the cause of your observation.

Best regards.

PS. I think this protein is an example of a so-called amphitropic protein, see for more info (and more info about the use of SOPMA, Heliquest etc.):

Veijo T Salo

thanks for answer. I might try this approach.

My guess would be that the fusion protein is unstable and is the target of proteases that resist the additions of EDTA and AEBSF.

My first choice protease inhibitor always is PMSF. Made freshly on the day of use in a water-free solvent.

And E. coli expression strains with some of the protease genes deleted also offer an advantage.

Many times the problems are in the linkers between MBP and your protein. Of course, linkers are designed to be flexible and allow independent folding of the two parts, however, they also are unstructured and thus easily targeted by proteases. Try to play around with linker length, very likely a shorter linker will solve your problem.

Do you see the full-length protein when you dissolve some cells in denaturing buffer (containing urea or SDS) and load them on a gel?

check this video , it may be helpful about GST fusion protein

Hello Sudharma Banerjee

The following could be the reasons.

1. High salt concentration in the samples could result in increased conductivity, affect protein migration, and can result in protein bands spreading into adjacent lanes. Remove salts from sample by dialysis or desalting column prior to sample preparation.

2. The ionic strength of your sample may be lower than that of the gel, causing the samples to diffuse out of the lanes. You need to recheck the buffer composition.

3. Sometimes, uneven gel interface could also be the cause. Overlay gels carefully.

Hope this helps.

Good Luck.

It doesn't seem that far off to me. Without knowing anything else I would suspect this is just do to random variation in your data.

Dear Suwannee Ganguillet ,

I first recommend you to read the following paper: Jass, J., Tjärnhage, T., & Puu, G. (2000). From liposomes to supported, planar bilayer structures on hydrophilic and hydrophobic surfaces: an atomic force microscopy study. Biophysical journal, 79(6), 3153-3163.

I noticed that you try to use DOPC. This is a typical bilayer forming lipid according to the structure-shape concept, see for example:

Cullis, P. T., & Kruijff, B. D. (1979). Lipid polymorphism and the functional roles of lipids in biological membranes. Biochimica et Biophysica Acta (BBA)-Reviews on Biomembranes, 559(4), 399-420. (free in Google Scholar, see figure 7)

Seddon, J. M. (1990). Structure of the inverted hexagonal (HII) phase, and non-lamellar phase transitions of lipids. Biochimica et Biophysica Acta (BBA)-Reviews on Biomembranes, 1031(1), 1-69. (see enclosed file)

My estimate would be that DOPC might be too stable, and you need a certain portion of so-called non-bilayer forming lipid.

Furthermore, in principle PC is considered as a neutral lipid, perhaps you need a certain portion of anionic (negatively charged) lipid.

My suggestion would be to first try a, in the literature, well-described mixture (in terms of composition and method) to make that work in your hands as well and then start to vary.

Hope this helps you somewhat.

Best regards.

Hi,

The most common palladium oxide is PdO with palladium in +2 oxidation state.

Palladium reacts directly with oxygen above 800 ^oC form PdO as black powder. In your case, it is probably the same. Mild oxidizing agents can also convert PdCl₂ or K₂PdCl₄ to PdO.

Best

Balakrishna

For predictive simulation of these processes, we use Autodesk Netabb (https://www.autodesk.de/products/netfabb/features), which works great but is cost-intensive. We also use the Simulia Abaqus "AM Modeler" Plugin (https://info.simuleon.com/blog/using-abaqus-to-simulate-additive-manufacturing-printing-an-optimized-hip-implant) which costs less and also delivers good results for this process. These simulations might potentially help you to reduce the thermal and stress-induced defects and deformations in your parts. Further, it might support you to improve your support structure placement (if required for your process).

When considering micro-defects and the quality of the material after manufacturing, you should specify the type of defects that should be "removed". Maybe a simulation would not be appropriate and you should consider an In-Line process monitoring and control for your process, combined with a proper "Design of Experiments". The data achieved by this procedure could be used to improve the simulations mentioned above by considering your materials and machines constraints.

Thank you Dr. Manuele Martinelli. I will try to locate the problem.

To do this properly, you need the reference enthalpies and heat capacities of the solid, liquid, and gas forms. I suspect you know this and do not have this information. If you know the melting and boiling points, you can estimate the reference values from a Gibbs Energy analysis (melting point is when the Gibbs energy of the solid and liquid are the same and less than that of the vapor and boiling point is where the Gibbs Energy of the liquid and gas are the same and less than that of the solid). However, you cannot estimate the reference enthalpy and entropies for all three phases (you need both for a Gibbs energy analysis) from just two data points. There are group contribution theories and some quantum-chemistry-based partition function approaches to estimating these if you know the structure well enough.

Malcolm Nobre , I don't see how freezing protein droplets in liquid nitrogen makes sense. You need to store the protein protein in a tube to put it into the minus 80 freezer. So what's the point? What's innovative about writing a publication about flash freezing proteins in tubes in liquid nitrogen? Doesn't everyone do that!?

I use FiJi open source software addin to ImageJ. You can load all the images to stacks in there easily and automatically.

Have you run a negative control of your non transformed cells? If they are present also in the negative control lane, both bands must be unspecific binding or your antibodies.

Make sure that your sample is heated evenly when boiling prior loading and always heat again before loading the gel even if the sample has been boiled before and stored at -20ºC.

I have seen several bands of the same polypeptide when my samples has not been heated evenly, this happens when some extract is stuck under the cap of the tube while reheating and then, after spinning down the sample just before loading, the boiled sample at the bottom of the tube is mixed with some extract that has not been heated from the cap, producing bands that migrates differently in the gel despite being the same polypeptide.

Hi dear Mustafa Nile

Did you observe the following steps?

Dr Md Moshiur Rahman That is an excellent & debatable question. These are the principles I try to follow in making this decision:

Other than obvious instability, all others are relative indications for fusion. One has to realize that fusion is a bigger surgery (more blood loss, operative time, length of stay, postop rehab), and has risks of adjacent segment degeneration, instrumentation-related complications and reoperations. It is a non-physiological remedy, and certain occupations may have restrictions for returning to work after fusion surgery. Lastly, recurrent discectomy is a reasonable choice for the first recurrence (with absence of instability). But second recurrences should likely be fused. Hope this helps. Thanks

Multiple options here: You could use traditional approaches from ensemble learning (stacking, boosting, etc.) to combine separate models for text and images. Alternatively, you could combine multiple sources in one neural network by concatenating fixed-size embeddings for text and image. For text, the easiest way would likely be the hidden state of a RNN or LSTM. You get a lower-dimensional representation for images after multiple convolution/pooling steps in a CNN.

That being said, the first option is likely going to work at least somewhat reasonably if any either text or images would be enough by itself to train a model. On the other hand, the second option is "mathematically sound" but might require prohibitive amounts of computing.

Stuart Russell's "AI a Modern Approach" and Aurelien Géron's "Hands-On Machine Learning" describe ensemble methods, and François Chollet's book on Keras outlines how to combine different data sources in one NN.

I hope that helps.

There are two basic EDS methods, electron-based EDS (usually electron microscopes-based) and X-ray based EDS (XRF). Electron based EDS is most sensitive to the lighter elements, while XRF is most sensitive to heavier elements. Both methods can detect all elements if they are in high enough concentrations. The neutral point is usually some where around P to Cl, depending upon the total electron voltage of your probes. Both methods can be quantitatively capable if well calibrated with an appropriate standard Containing all involved elements in detectable amounts. You can create your own standard as long as each involved element is present in detectable amounts. At least 6 independent calibration analyses are needed of your standard for the calibration, and all 6 of these analyses need to be averaged together and then entered into the system to be used as your standard. I would recommend at least 6 analyses of your experimental material as well.

Dear Prajna Deepthi many thanks for posting this interesting technical question. As an inorganic chemist I'm certainly not a proven expert in the field of polymer chemistry. However, I would not be surprised when certain physical properties of PTFE polymer change in one or the other direction when you add non-metallic graphite on one hand and graphite coated with metallic and highly conductive copper on the other hand. For some more information about this please have a look at the following potentially useful articles:

This paper has been published Open Access and can be freely downloaded as pdf file from the internet (see attachment)

(also attached as pdf file)

I hope this is useful. Good luck with your work and best wishes, Frank Edelmann