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Biochemistry - Science topic
This group is designed for the exchange of thoughts on Biochemistry
Questions related to Biochemistry
Dr. Faria is associated with Department of Biochemistry (U38-FCT), Faculty of Medicine, University of Porto , 4200-319 Porto, Portugal.
📢📢📢 Call for Papers: Memorial Issue to Prof. Kazuo Umezawa: A Noteworthy Biochemistry Educator
📅📅 Submission Deadline: 31 December 2024
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Welcome your comments~
Hello!
I was wondering if anyone had a detailed protocol for measuring β-hexosaminidase activity in isolated lysosome fractions.
I will have 0.5ml fractions of lysosome isolate (after magnetic isolation).
Here is what I know so far:
- samples: lysosome aliquots with and without Triton X-100 (1% final triton concentration) + whole cell lysates + cell culture supernatant + controls without sample
- reaction buffer: 100 mM Na Citrat, 0.2% BSA, 1mM 4-MU-β-N-acetylglucosaminide
- incubation: 10 min 37°C
- stop solution: glycine/NaOH pH 10.4
- fluorimeter: excitation 360nm, emission: 450nm
Serán empleados como antecedentes de una investigación con este propósito
I have to estimate the growth of my bacterial cultures spectrophotometrically and I read articles of measurement at an OD of 600nm. Also what to do if the values exceed 1. What is the proper method for the measurement of the same.
Cure of cancer?, respectfully to whom it may concern:
If I found a cure of cancer, I’m wondering what I should do with it? I’m just coming out of a 40 days fast of only drinking maple syrup and pure lemon juice plus a little cinnamon powder and nothing else since I had unimaginable pain in my colon end. I figured out that although we humans can last for weeks on this diet /fast, cancer has no food on that and just goes away and the pain is gone too. I received this prescription by tuning in into the HeilstrOm and I figured out that it also enhances the reception of the HeilstrOm greatly, which brings great happiness too, as I describe in my book
Book yogapsychologie
Best regards, hope this helps someone
Chris K. Frueh
Independent Researcher
In the identified case of familial desminopathy (T341P DES mutation in heterozygous state), the son has bradycardia, but the father did not have bradycardia. How can this fact be explained?
I am measuring the specific activity of an enzyme under two different conditions using an enzymatic kinetic assay.
In one of my conditions, I know that the expression of the enzyme is highly upregulated. Therefore, even though I use equal amounts of sample (protein concentrations measured by BCA), I expect higher levels of the enzyme of interest in this condition.
In case I am investigating the activity per molecule of enzyme.
So, would it be appropriate to normalize the specific activity by dividing it by the enzyme expression data from a Western blot?
*Note: I subtract the background activity (before substrate addition) and non-specific activity (after addition of a specific inhibitor) to calculate the specific enzyme activity.
Dear researchers,
i would like to make a research on "high quality organic fertilizer production from biogas extracts"
looking forward to your sugessions
I was wondering if it is possible to form a permanent open "ssDNA bubble" similar to a transcription bubble (>13 nucleotides) within E. coli. These criteria are important:
1. Open ssDNA bubble within replicable (in E. coli) genetic element. So no C-Traps under force.
2. No proteins, nucleic acids, or other toxic chemicals supporting the bubble. Can help during nucleation, but bubble has to be accessible for protein interaction.
3. Stable in bioorthogonal conditions. Physiological pH, salt, 37 °C, etc.
I was performing estimation of Total Soluble Sugar content in bacterial cultures by Anthrone method. Out of 9 samples, only one sample showed positive reading while others had a negative reading in the UV-vis spectrophotometer. What does this denote.
In My article: The Extended Pedersen hypothesis (published, February 1988 Clinical Physiology and Biochemistry 6(2):68-73,) there is mistake of the name of first Author: it is not Catherine Macfarlane, but Campbell M. Macfarlane.
Can you please correct it?
I have extracted essential oil from a medicinal plant and want to make several concentrations for some biological activities (Antidiabetic assay). How can i prepare the several concentrations to determine the IC50.
What will be the solvent for the dilution of the essential oils?
Your kind help in this regard will be highly acknowledged.
It is known that patients with desminopathy often die from pneumonia. Have pathomorphological studies of the lungs been performed in patients with desminopathy?
Hi! I'm doing a renilla luciferase assay with coelenterazine and HEK293T cells transfected with AP-1 renilla luciferase. I'm wondering if the cells need to be lysed prior to adding coelenterazine and measuring the luminescence. Coelenterazine is cell-permeable, so I'm wondering if/why the lysis step would be necessary.
I understand that the proteins would be more exposed--would the luminescence be not as great through the cell membrane if we didn't do the lysis?
We are researching the conjugation of various antibodies to quantum dot microspheres and europium microspheres. The sustainability results have been unsatisfactory. Which stabilizing buffers do you recommend for stabilizing the conjugation step?
Best regards
I have purchased Acetylcholiesterase from Electrophus electricus (electric eel), C3389-2KU, following details are written on the enzyme vial.
Type-VI S lyophilized powder 200-1000 Units/mg protein, 374 units/mg solid, 610 Units/mg protein, 5.3 mg/solid.
i want to prepare 0.28 U/ml working solution, please help me with the preparation of stock solutions and the calculations.
What is the difference between absorption and adsorption?
Hi, are there other methods beside Western Blotting to detect a specific protein? I‘ve isolated mitochondria and want to show that hPar17/Pin4 is present in mitochondria. The Western Blots didn’t work well, because of the low amount of sample or maybe low expression levels of hPar17 (the Abs work). I want to detect the protein without overexpressing it if possible. I isolate the mitochondria from human cardiac myocytes. Do you have any ideas on how to show the presence/expression of my protein in mitochondria? Thank you very much!
Hi, I am looking to perform an analysis of Br/ KBr in samples of water sources to see if the levels are safe. Due to budget and resource limitation at the stage, I would prefer a method that considers these factors.
Kindly help.
I am planning to purify the mononucleosome for cryoEM study. I have seen, people assemble the nucleosome from recombinantly expressed histones and then providing DNA sequence to it. Isn't this possible to islolate nucleosome and after enzymatic cleavage, followed by SEC chromatography for the cryoEM study ? I understand that there could be a population of others but it could be more relevant. Please suggest.
Hello,
My lab recently acquired a combination ORP electrode, with the refrence being Ag/AgCl and the working electrode being platinum.
We are hoping to use this electrode to measure the reduction potential of buffers prepared using biologically active compounds, such as GSH and GSSG and probe redox active systems by artificially setting the potential.
Recently, I was trying to validate the electrode by preparing 1 mM total concentration solutions of varying ratios of GSH and GSSG. This was done as, to my understanding regardless of concentration the ratio of Ox vs Red determines the potential value of the solution via the Nernst Equation. However the readings I got were all positive, and nowhere close to the expected potential, even when correcting for the electrode difference between Ag/AgCl and SHE.
Secondly, in 1x PBS pH 7.4, I added increasing amounts of BME up to 1 M and got an exponential decay like curve asymptotically approaching ~-120 (SHE) mV.
I am having trouble making sense of these results, namely the GSH vs GSSG ratio, and why the readings would not follow the nersnt equation.
Can anyone explain how to use these ORP electrodes, and where I may be going wrong in these experiments? All the information I can find online are referring to waste water treatment.
I am attempting to synthesize a schiff base ligand by reacting 5-hydroxytryptamine (HCl salt) and pyridoxal (HCl salt) in methanol, but the final product is not crystallizing in the methanol to be recrystallized. The product is originally a sticky brown oil that solidifies as a whole. What should I try differently? Sulfuric acid catalyst and maybe glacial acetic acid? Anything else?
I tried glycine buffer but the yeast cells acidify the buffer so the pH goes down to 7-6 overnight. I need something that will stay around 9 and that isn't toxic to the cells.
Can a published journal article be submitted to conferences?
Most of the researchers use to teach at university. In some careers, professionals who exert their profession without doing research share teaching spaces. When I was a chemistry student, 100% of my teachers were researchers ranging from PhD candidates to experts in their respective fields. While it may seem logical for researchers to be the best candidates to teach in fields such as chemistry or biology, what about healthcare-related fields like medicine, pharmacy, or biochemistry? Who is better suited to lead a class, a researcher or a professional, or both, each one in different subjects? We can distinguish between basic and clinical subjects. I am interested in hearing your thoughts on this matter.
In all academic sources, sucrose is identified as α−glucose (1-->2) β−fructose. However, I cannot find any explanation anywhere as to why the fructose monomer has to be in the β configuration. Maltose has both α and β anomers, same for lactose. Even trehalose, another non-reducing disaccharide with glycosidic linkage between two anomeric carbons, has α-α, α-β, and even β-β anomers. Why is sucrose special? And is there a disaccharide out there that has α−glucose (1-->2) α−fructose configuration?
A patient with desminopathy (mutation Thr341Pro DES in a heterozygous state) with the progression of the disease has a decrease in taste and smell, immunosuppression, and an increase in IgA in the blood.
Oddly enough, but all this is characteristic of infections, including viral ones. For example, it is known that if the hepatitis C virus is not treated, then death will occur in 20 years.
In the identified case of late onset desminopathy, muscle weakness manifests itself at the age of 30, and death occurs 20 years after the onset of the disease.
Could the desmin mutation in myofibrillar myopathy be caused by an infection?
Perhaps the infection contributes to the progression of desminopathy?
I need some suggestions for articles recently focused on the computational design of proteins, as well as the evaluation of the designed proteins by assessing various properties or values. If anyone is interested in this field and has read articles on this topic, please don't hesitate to share your suggestions below
Dear Researcher Gate,
I inquired about adding a reviewer to the submitted manuscript in Biochemistry can you help me with this?
Thanks
maha saad
I am currently learning about PyMol to utilize in my project. I used PyMol to visualize potential H-bond interactions in specific amino acid residues. However, I have discovered that Arg465 and Ser461 show a distinct interaction, as shown.
Please help identify this interaction.
An unopened Sigma-Aldrich (P4557) phenol solution bottle was shaken (prior to the addition of the Equilibration Buffer) and a gel-like layer formed at the bottom of the bottle. The upper phase is still liquid. The bottle was shaken briefly after the phenol solution was taken out of +4 C. What should be done? Should it be heated in order for it to return to liquid?
Hi,
I am looking for build up a consensus or a group in clinical biochemistry for mutual exchange in scientific idea, research protocols, cooperation in proposal preparation, sharing in books, Research articles and reviews.
It is known that in the early stages of desminopathy the muscles most often affected are: Semitendinosus, Gracilis and Sartorius. What is the reason for the damage to these particular muscles?
In biochemistry, most work is done at room temperature. Yet, basic thermodynamics tells us that affinities often change with temperature, in positive or negative directions depending on the entropy/enthalpy contributions. Thermal transitions can occur which both quantitatively and qualitatively change the behavior of the molecules of study. In enzymatic reactions rate-limited by diffusion-mediated product release, the increased rate of diffusion could increase the rate of product release above the rate of the chemical step, such that the chemical step becomes rate-limiting. If one discovers a compound that potently inhibits this enzyme at 25C, it may have reduced effect at 37C, or none at all. Likewise, if an enzyme is predominantly dimerized at 25C based mostly on enthalpic contributions, this dimer may not even exist at 37C. Screening compounds against the dimer may be of little relevance to the situation in vivo. The converse could happen if dimerization is entropically driven. Temperature-dependent changes in solution properties can also obscure the relevance of 25C results to 37C, such as viscosity.
I welcome everyone's two cents.
Here is an excerpt from Kollmann et al. (2020, J. Physiol.):
"This ring was placed in a recording chamber continuously perfused with 37°C aerated Hepes solution containing (in mM) 136 NaCl, 10 glucose, 5 KCl, 10 Hepes, 1.2 MgCl2, 2.5 CaCl2 (pH 7.40) at a rate of 11 ml min-1. Hypoosmolality of the Hepes solution was established by reduction of the NaCl content to 33 mM (94 mOsm kg-1 H2O), 58 mM (144 mOsm kg-1 H2O) or 83 mM (194 mOsm kg-1 H2O)."
For validation, I tried to calculate the osmolality by myself. For example, when the NaCl content is changed to 33 mM, the osmolality should be
33*2 (NaCl)+10*1 (Glucose)+5*2 (KCl)+10*1 (Hepes)+1.2*3(MgCl2)+2.5*3(CaCl2)
which gives 107.1 mOsm/L, i.e., 107.1 mOsm/kg. But 107.1 is largely different from what the author stated, namely 94 mOsm/kg.
When the NaCl content is changed to 58 mM or 83 mM, the osmolality should increase by (58-33)*2=50 mM, or (83-33)*2=100 mM. This is consistent with the author's calculation, i.e., 144-94=50 mM, or 194-94=100 mM. So, my calculation is correct at least in terms of NaCl. But how can I calculate the contributions of the remaining solutes correctly?
Reference: Kollmann, P. et al. Submucosal enteric neurons of the cavine distal colon are sensitive to hypoosmolar stimuli. J. Physiol. 598, 5317–5332 (2020).
Is it correct to filter the extracts we obtain with Whatman filter paper and use them directly in experiments to investigate the bioactivity of phytochemical substances? Or is it correct to first centrifuge the extracts we obtain, then filter the supernatant and use them in experiments?
Some (but not all) DNA polymerases such as Klenow fragment, Taq, and Phi 29 DNA polymerase can catalyze strand displacement synthesis. This is evidenced by opening of molecular beacons and Loop Mediated Isothermal amplification (LAMP).
in strand displacement synthesis, the primer strand is extended using the template strand and each nucleotide incorporated displaces the 3rd strand that was originally annealed to the template?
where is the third DNA strand? the one being displaced? crystal structures and cryo-EM structures of linear primer template dsDNA structures with magnesium and dNTPs and DNA polymerases are not informative about where 3rd strand of DNA is. The displaced strand is not the template and is not the primer.
Where is the displaced 3rd strand of DNA?
I need to dissolve sulphate to purify the nanozyme synthesized with magnesium sulphate
- Which is your favourite field of Medicine and why?
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- What is your field of interest?
Dear all,
I have been trying to knockdown my target protein using siRNA. The protein has several isoforms the effect of this knockdown has been showing up in the functional assay. However, I don't see the knockdown in the wesptern blot of the same test sample. Please suggest how to do an effective knockdown so that I can visualize that in the western blot as well.
Thank you in advance for your kind suggestions
Sincerely,
Prem
Hello,
We are trying to purify proteins using a secretion system but do not have TFF cartridges for our device. Does anyone know where we can purchase these? Or is there a better replacement? We want to downscale the volume from 3 L to 100-200 mL.
I've attached a picture for reference.
Thanks,
Thomas Newton
I'm on the lookout for remote bioinformatics and computational biology opportunities where I can actively contribute to research projects. Compensation is not a priority for me; my main focus is to gain hands-on experience in these fields.
#biopython
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I intend to explore the integration of computational biology and artificial intelligence (AI) with laboratory and experimental work, encompassing animal models, cell culture, clinical trials, and molecular studies. As a clinical biochemistry student with a keen interest in AI, I believe this interdisciplinary approach holds immense potential for advancement and innovation.
However, I face the challenge of identifying relevant literature in this emerging field. I would greatly appreciate guidance on effective keywords and search strategies to navigate this landscape of research and achieve my research goals.
My target protein is a membrane protein and I want to check its expression level in the test sample using western blot. Can anyone suggest which protein should be used for loading control, like we use beta-actin in cytosolic proteins. And is there antibody available for that ?
I intend to explore the integration of computational biology and artificial intelligence (AI) with laboratory and experimental work, encompassing animal models, cell culture, clinical trials, and molecular studies. As a clinical biochemistry student with a keen interest in AI, I believe this interdisciplinary approach holds immense potential for advancement and innovation.
However, I face the challenge of identifying relevant literature in this emerging field. I would greatly appreciate guidance on effective keywords and search strategies to navigate this landscape of research and achieve my research goals.
I need to convert from micromoles per second per liter (µmol s⁻¹ L⁻¹) to millimoles per gram of dry cell weight per hour (mmol gDCW-1h-1)
Thanks!
Hello everyone! I am new to this platform, I am looking for this book: "Bioquimica del estres Oxidativo" (Diego Camps, 2010)
I would like to know if anyone has it in digital format to share it with me. Waiting for a reply. Thank you very much!
I need to extract the liver from frozen Emerald rockcod and sequence its RNA. To keep the RNA stable and prevent degradation, I would like to avoid thawing the fish for the dissection. However, I haven't been able to find any methods that keep the fish frozen. Does anyone have any tips on how to best achieve this?
If an active site mutant knocks out product formation at all excessive concentrations of substrate and at all excessive concentrations enzyme, but substrate binding affinity via anisotropy shows no difference in binding affinity for wildtype versus active site mutant enzyme, is kcat = 0? But if kcat = 0, then by the relationship of Michaelis constant (KM) to kcat then KM = KD.
How do you show to reviewers that the active site mutant is dead? If the wildtype mutant starts producing product in seconds, are you supposed to measure the reaction for the mutant for an hour at zero concentration of substrate and at 100fold excess substrate concentration of the K_M for the wildtype enzyme for the active site mutant, and show the time course?
Are there any publications that show a mutant enzyme is not just slow to produce product but is rather incapable of producing product but can still bind substrate? Examples would be greatly appreciated!
Hello all,
I hope everyone is doing well.
We are preparing buttermilk from standardized milk. In this regard we need to increase the pH of the buttermilk. We used sodium citrate (0.4% & 4%) and sorbitol (0.4%) to check any change in the pH. Unfortunately, there were no significant changes observed.
Please suggest any recommendation to increase pH using any natural or food-grade chemical compounds and any other alternative options.
Thank you in advance.
Keep smiling and Stay healthy.
I am reaching out to #researchers in the field of #Biochemistry, #Biophysics and #Bioinformatics, for collaborative partnership in scientific research. The researcher should be academic staff at the tertiary institutions in following listed countries:
#Afghanistan
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#Burkina Faso
#Burma
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#Congo
#CookIslands
#Cuba
#Democratic People's Republic of Korea
#Democratic Republic of the Congo
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#Dominica
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#Eritrea
#Eswatini
#Ethiopia
#Gambia
#Ghana
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#Guinea
#Guinea-Bissau
#Guyana
#Haiti
#Iran
#IvoryCoast
#Kenya
#Kiribati
#Kyrgyzstan
#Lao People's Democratic Republic
#Lebanon
#Lesotho
#Liberia
#Madagascar
#Malawi
#Maldives
#Mali
#Marshall Islands
#Mauritania
#Micronesia (Federated States of)
#Mozambique
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#Niger
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#Moldova (Republic of)
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#Senegal
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Interested researcher should kindly email to hezesapience@gmail.com with the subject: Research Collaboration from "your country".
Thanks.
Toluwase H. Fatoki
Visionary @ Heze-Sapience International, Nigeria.
Lecturer @ Department of Biochemistry, Federal University Oye-Ekiti, Nigeria.
Is there any system with small molecule binders and a short protein tag that is higher affinity than 6xHIS-NTA?
Dear all, please suggest or provide a link for the provider or company who can design or synthesize shRNA plasmid (Single vector system) that will directly express into the mammalian cells to knockdown the protein expression for long term. Any suggestions will be highly appreciated.
Thank you
with kind regards
Prem
I am trying to express several proteins at the same time, but I want to use a different promoter and terminator for each one to avoid the possibility of recombination.
The promoters that are available to me are: TEF2, PGK1, CCW2, TDH3 and HHF2. The available terminators are: ENO1, SSA1, ADH1, PGK1 and ENO2.
Has anyone ever used these combinations of promoters and terminators? In your experience, which combinations work the best?
Hi
I'm purifying some mutants of the protein I study. The wild type protein exists as a monomer and is 28kDa.
I have two mutants (same protein, same number of amino acids but with 8 amino acid substitutions at defined positions), one of the mutants (mutant 1) analysed using size exclusion chromatography with multi-angle static light scattering (SEC-MALS) and its MW was shown to be 33kDa and has an oligomerization state of 1.2. The other mutant, mutant 2, also measured by SEC-MALS was 59kDa with an oligomerization state of 2.2.
For the wild type to measure the concentration I've just been using the MW (28kDa) and extinction coefficient (calculated by entering the sequence into online software ProtParam) and using a NanoDrop measuring absorbance at 280. This gives the concentration in mg/ml which I then convert to molar concentration.
For the mutants I want to measure their concentration the same way - measuring A280 on the NanoDrop using the mutants MW and extinction coefficient and calculating molarity from mg/ml. I'm not sure if this is an obvious/stupid question but what MW weight and extinction coefficient would you use for the mutants on the NanoDrop? E.g. For example mutant 2 molecular weight (MW) of the protein based on its amino acids (AA) composition is predicted to be 28kDa, but SEC-MALS shows it is 59kDa as the protein forms an oligomer.
My instant is to use 59kDa and the computed extinction coefficient predicted from the AA composition - is this correct?
Thanks in advance!
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I learned DNS assay to find out the quantity of maltose in the sample.
I realized the importance about the boiling water in this assay. But I got a several question after that.
I don't know the reason why do I have to cool-down the sample after heating up?
And also is ice a tool used to reduce cooling time, or is it used to cool quickly?
Please leave me a response about this problem.
Thank you for your help.
Joonseo_Cha The Student of Hallym University
The size differ little bit from one species to another, yet they have one size range. Also, the size of them in their native form so they don't lose their colour while isolation.
I saw these on a same step of conversion of Malonyl CoA to Malonyl-ACP. Could anyone clarify this? I add that reference link below.
KEGG PATHWAY: Fatty acid biosynthesis - Chlorella variabilis (genome.jp)
I'd like to perform a SEC step prior to on-column refolding of a protein I am expressing/purifying but am worried about the extended time the protein will be in the presence of urea (and subsequent protein carbamylation) in the current refolding workflow I have.
Is there any concern of protein modification with a 4-6 hour denatured protein purification workflow using 6M GndHCl?
I am a second year PhD student at Wayne State University looking to screen a library of photosensitizers (transition metal complexes) for activity with ultrasound irradiation in cell cultures (likely MCF-7A cells). I would ideally like to set this up in a 96-well plate to screen multiple compounds but I am unsure of the set up as previous literature has not been as helpful as I would hope. The instrument I have is 3 MHz with a 3.5 cm diameter probe (Model-Sonic 103, Preamonex). I am unfamiliar with soundwaves so I will briefly discuss what I know so far from previous literature.
Overall, 0.3 W cm–2, 3.0 MHz seems to be a fairly common treatment condition when combining compounds with cell cultures.
This source clusters samples in a 2x2 square in the well plate and places the probe underneath the plate per cluster (see SI). Does a coupling agent need to be used and can this be done while keeping the plate sterile for continued incubation? Do the ultrasound waves pass through the plate to other clusters of wells (in a way that would significantly affect the other clusters)? Is there a way to evenly apply ultrasound stimulation evenly across the whole plate?
This source works with nanoparticles and a 96-well plate, but details of the set up are not given. Unless I'm missing something?
This source appears to place a sponge in degassed water on the head of the 35 mm probe. Could this be a valid option for the cluster of wells (2x2 square) mentioned above?
For the purpose of transfection, a 6-well plate was put into an ultrasound water bath.
Ideally, I would like to use the probe (as opposed to a water bath) to keep the instrumentation consistent between in vitro and in vivo studies. As I mentioned, I'm very unfamiliar with sonication/ultrasound and its physics. Any assistance/advice on how to make an accurate and precise high throughput set up is appreciated.
Thank you in advance!
How often do you get new salts? What salts do you get "fresh" most frequently? All help is greatly appreciated. Thank you to everyone who reads or responds, hope you all have a fantastic day.
I am a Graduate Student at the University of Wisconsin Milwaukee. Our most recent electrophysiology experiment was going fantastic for about 3 weeks. Suddenly, every slice was very poor quality, and every cell we patched onto died within about two minutes. Even if you don't have an Electrophysiology background, all advice and input is appreciated. It's been two weeks since this problem first arose. We didn't alter any procedures, everything has been held constant. We don't know what could be causing the sudden change in slice quality.
So far we have three theories: DDH20 filter needs replacing, our glassware has somehow become contaminated, or our salts have gone bad.
The DDH2O water resistance reads about 14.8MOhms. We changed the filter about a year ago. From what I have read, DDH2O should be between 14 and 18 MOhms to ensure slice quality, so we think our water is fine.
Our glassware washing procedure is 3 rinses of tap water, 3 rinses of deionized water, and 1 rinse of DDH2O. We don't know how else we can safely clean the glassware for slice preparation. But we don't think this is the issue.
Our main theory is the salts have gone bad. Our lab is on the 4th floor, and it is usually quite warm and humid on our floor during the summer. Most of opened salt bottles we use were first opened between 2 and 7 years ago. (for example, our bottle of sodium phosphate monobasic monohydrate and our potassium chloride were both opened 6 years ago. Our magnesium chloride and sodium chloride were both opened 2 years ago)
We think with repeated opening of the salt bottles causes debris/water from the atmosphere to leach into the salts, resulting in a change in our solutions and causing the cell death we have been observing. Is this a real possibility? How often do you get new salts? What salts do you get "fresh" most frequently?
We checked the pH, the pH of our solutions is within .1 of what it should be. All help is greatly appreciated. Thank you to everyone who reads or responds, hope you all have a fantastic day.
I am currently making bacmid and consequent baculovirus using pDEST8, pFastBac and 438a vectors and recently switched to DH10EmBacY cells instead of DH10Bac due to the added YFP signal to monitor transfection, etc.
I was wondering if there is any reason for me to not use either of the two competent cells interchangeably to make bacmid DNA?
I observed an interesting pattern that resembles a "flattening" of the near-UV region of the absorbance spectrum of the rat intestinal mucus in the streptozotocin-induced model of sporadic Alzheimer's disease. I'm studying qualitative and quantitative alterations in mucus as there seem to be some changes in the gut-brain axis, intestinal redox homeostasis, and cell turnover ( , ). I observed a quite dramatic change in the number and responsiveness of goblet cells so the observed spectral shift may be driven by a component of mucus. Another possibility may be a component of bile, a peptide or bilirubin/biliverdin, or even complexation between bilirubin/biliverdin and some other molecule (e.g. I found a similar pattern in a publication by Klinke et al. who reported flattening of the biliverdin near-UV spectrum upon binding to bacteriophytochrome? ). Any ideas about what I may be looking at?
Thanks!
Dear community,
I am planning on conducting experiments for which I need to obtain a plasma-free platelet suspension from an aliquote of a platelet concentrate. Do you now any methodology/protocol that allows for washing without extensive cell activation? I need the cells to be a "functional" as possible.
Yours sincerely,
Michael
I would like to add BG-azide to cells for SNAP tag pulldown however I don't know whether it is going to be cell permeable. My instinct is to day yes it will be but neither me nor the supplier know whether that is true. I thought I would ask if anyone has tried something similar, even though that is unlikely... I have attached the structures in case that is informative
These are the products https://www.iris-biotech.de/global/rl-3950 and https://www.iris-biotech.de/global/rl-3960
In a patient with desminopathy (mutation Thr341Pro DES in the heterozygous state) with the progression of the disease, we note signs and symptoms that are also characteristic of botulism: bradycardia, arrhythmia, AV blockade, a significant decrease in the average duration of motor unit potentials according to electroneuromyography, paresis and paralysis of the striated muscles, decreased sweating, paresis of the gastrointestinal tract, dry eyes, dry mouth, symmetry of neurological symptoms, hoarseness, impaired visual acuity, doubling of objects occurs, progressive muscle weakness. These signs and symptoms are characteristic of botulism, only when a case of desminopathy is detected, they proceed slowly.