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Can we use Agarose gel instead of PAGE for protein sepration?
Because agarose gel is easy to make
no. Please refer some basic molecular biology book.Following
Does anyone have experience using infrared secondary antibodies for western? What protocols work best for you?
I am trying to detect a purified bacterial his-tagged protein on western blot (using nitrocellulose membrane). I did see the protein in elution fractions on SDS gel using coomassie stain. Primary used to detect is a rabbit anti-his, and my secondary is anti rabbit-680 conjugate. I blocked with 5% milk, incubated with primary o/n at 4 in 5% milk, and also placed the secondary in 5% milk for 45min at RT (I was told by some protein loving friends that milk can also help improve secondary antibody background). I have not used these fluorescent antibodies before. Called the company and am waiting to hear back, however in short, it might be that I should tailor my staining protocol to something closer to an IF, so use serums for blocking and incubations instead of milk. Also an option to just use TBST for antibody incubations. Any help/thoughts is much appreciated!
we are using an Odyssey system for performing WBs with fluorescently labelled antibodies. Here, LI-COR provides a specific blocking reagent which we use routinely and obtain good results with. I also use this reagent to dilute the antibodies. See link below
Hope this might help you.Following
SDS-PAGE - loading dye migrating strangely on precast gel
I am using Bio-Rad StainFree AnykD precast gels, and in many of them (I'd say four out of six of the box I have run) the dye has migrated weirdly - instead of being in a nice blue hazy band it looks as though there are two blue bands - one in the gel, and on almost in front of the gel (but behind the plastic). This also happened to another lab member once who was using a different box of gels, different apparatus and buffers.
I feel like it might be the speed of the run? 200V for 30 min. We use Tris Glycine buffer. Has this happened to anyone before? Any idea of what causes it?Following
Should university teachers have experience in Primary Education before teaching at the university in education degree programmes?
I am so interested in your opinions about how to teach at the university in different countries. Is experience needed? Can you teach without experience in the field you will teach?
This is an interesting question. Our research is showing that those teachers who had unremarkable educational experiences, who stood outside the norms of teaching and learning make the most effective teachers. So perhaps it has less to do with whether the teacher taught primary school and more important that the teacher is informed, critical of unexamined practices and able to see possibilities for effective educational change.Following
Can any body inform me about the cross-roads between innate and adaptive immunity in the oral cavity?
oral cavity forms the mucosal immune compartment that belongs to the common mucosal immune system.
May be this article will be of your interest.
In general Chemokines and cytokines are considered such cross roads. You can refer below review of mine as well
What is power angle spectrum (PAS) and AS of the azimuth angle in 2d and 3d channel?
just explanation pleaseFollowing
how can i build research using Sentiment analysis on Patients shared text?
- which Data set is available contains Patients profiles,disease features, medicine and results , then the patient opinion on medicine or on doctor
- What will be my contribution point if i applied sentiment analysis on this data set.
I m not sure about the availability of the particular data, it will take extra effort to collect this such data.
The contribution point may be: This can be a feedback/review for the doctor or hospital, especially about their services.Following
Can anyone guide in synthesis of pyrrole boronic acid ester?
I am finding it difficult to synthesize pyrrole boronic acid ester? Does anyone have experience with boronic acid synthesis? Your suggestion might be helpful.
I have attached the reaction Scheme I am trying at the time. The desired product is not formed rather dehalogenated product is synthesized. I can't use lithation chemistry as there is ester.Following
Can we induce immunosuppression in mice using drugs such as ciclosporin A? What could be the efficiency of such induced immunosuppression?
We are planning to design platelet transfusion model in mice where we need immunodeficient mouse. But due to cost we are thinking of alternative strategy. If we could induce immunosuppression with drugs then it will be helpful.
Thank you Sandip Chakraborty for the answer.Following
Can anyone suggest literature on fault diagnosis?
i need some literature on fault tolerance systems and diagnosis.
i want to apply machine learning techniques .
Dear Khalaf i need your precious help regarding fault detection in railway suspension systemFollowing
How can I get rid of e-waste?
Up to 90 percent of the world's electronic waste, worth nearly US $19 billion, is illegally traded or dumped each year, according to a new report.
Use and throw - equipment manufacturer can make more money. So, they do not encourage repairing.
Size of the device is very small - miniaturized. Only machine can handle that.
Mass production of systems. Cheaper
The labor or service charge is high.
It is easier to identify the faults at board level and replace it than identifying the faulty component and replace it by de-soldering and re-soldering an SMD
So, the cost of repairing has become more than the new part. And customers want new product than refurbished.
So, it is unattractive to repair and use. So, we accumulate junk.
Recycle and reuse is the way to go.Following
Help required regaridng modeling of railway suspension system. i intend to design a simulink model.?
help required urgently.Following
Has anyone come across any recent research which has taken place on Academic writing teaching using social media?
If anybody has a clue on this area would welcome the feedback please.Following
Why Mg doping concentration cause increase in XRD peaks intensity of ferrites and chromites when compared with undoped sample?
intensities of peaks (111) (400) (422) (511) significantly increases with increasing the Mg doping concentration, why is so? any refrence to study please?Following
How can I assign a structure to "sys" in MATLAB s-function mdloutput function?
In the documentation it is mentioned that "sys" can be a column vector but I would like to output a structure from s-function block.
Ngoc check this:
What is the use of furosemide in AKI?
what is the use of furosemide in Acute Kidney Injury?Following
Please advise molecular visualization program for visualizing and calculating the intramolecular H-bonding ?
Free programs running under Windows, Thanks!Following
Arc length method for non linear buckling (Nonlin.-FEM in ANSYS): Is there any way I can continue the Simulation beyond the critical point?
I´m doing a non linear Simulation in which I have to deal with non linear buckling.
Are there methods or altered arc-length algorithms that can cope with the Issue of Stability (see picture)?
Thank you very much for your answers, especially Naveen. I appreciate it.Following
What does software as a service provide?
GMail is an example for SaaS, The s/w is provide, u can use it.Following
Can you suggest a measurement scale for behavioral intention to participate in research?
I am trying to locate recent studies which have validated and used scales for measure behavioral intention. Some papers report one-item measures like, Are you planning to xxx, or I intend to xxx. I am also trying to find studies that have focused on BI to participate in research studies (medical or otherwise).
Does anyone have any suggestions?
this is the website of Ajzen's theory of planned behaviors.
There, you will find all resources that you need - most notably a manuscript how to set up questionnaire items.
With regard to the number of item (of any construct): More is always better as you increase the reliability. The problem is that by slightly varying the content/question wording, you may measure different things. For instance, "I intent to do X" is not the same as "I plan to do X" - as planning is something that occurs after the individuals has decided to do X (i.e., the preparation). In the same manner "I will do X" is not intention but "self-prediction" and incorporates feasibility of conducting behavior beyond the mere intention (a reality check). Hence, with every measure, the devil is in the details.
In my career I have conducted thousands of CFAs with so called established mult-indicators scales and almost NONE of them fitted adequately the factor model. Hence, I vote for a very SMALL number but FOCUSED / clear items.
What are the differences between self-regulated learning and approaches to learning?
I am wondering, if i would like to look at how the way a course is taught can influence student's self-regulated learning, and their approaches to learning, should I seperate the two studies? Or they can be integrated? Can someone with the knowledge of what factors in learning and teaching influence self-regulated learning point me to some key literature?Following
What is the actual procedure to prepare QC samples in metabolomic profiling?
I am curious to know the actual procedure to prepare QC samples in metabolomics profiling. I would appreciate if you can shower your opinions on preparation of QC samples. Do we need to pool the biological sample for QC and extract the sample or can we pool the extracted samples from individual samples and prepare the QC. Your opinions or advise is much appreciated.
I think it really depends what you are trying to do when you run a pooled biological QC, as to whether or not you pool before or after extraction. If you are trying to capture all the variation in your procedure then it may be relevant to pool before extraction, but then I believe you are mainly testing the reproducibility of your sample preparation technique - and really this ought to be corrected for by the addition of internal standards. On the other hand if you are using the pooled biological QC to correct for the behaviour of metabolites in your analytical system, that are not able to be corrected for by the internal standard (i.e. metabolites that behave in an opposite pattern to the internal standard), then you really should be pooling after extraction and then split into individual aliquots that are run throughout your batch sequence at regular interval. Under these circumstances you have created a homogenous identical mixture that theoretically should give you identical chromatograms, but in fact will reflect any variations in your analytical system. These variations can be corrected on a metabolite by metabolite basis. For a really good paper on this you should look at:
The importance of experimental design and QC samples in large-scale and MS-driven untargeted metabolomic studies of humans
WB Dunn, ID Wilson, AW Nicholls, D Broadhurst
Bioanalysis 4 (18), 2249-2264
David Broadhurst has done a lot of work around the development of algorithms to make use of these pooled QC's. Minimally though, as suggested above use the QC's as a way to assess the quality of your data when performing a PCA. After transformation and normalisation they should be a tightly clustered group that sits into the middle of your PCA plot.Following
What is the relationship between ABA and ethylene on osmotic plant stress?
There are indications that ABA is a sign for stomatal closure on osmotic stress. For ethylene, an antagonist of ABA action. So if we block ethylene action, we would eventually favor ABA action. That was not what I found by using an ethylene inhibitor (silver), looking by the plant response: they became more susceptible to the osmotic stress.
We measured both phytohormones (ABA and ethylene). High level of these two phytohormones detected together with strong inhibition of stomatal opening.. I hope I help with discussion by this finding.Following
Where can I get a detailed analysis of hot water spectrum in the wavenumber range of 4000 to 400 cm(inverse)?
I am trying to study the IR spectrum of hot water in the waveno. range 4000 to 400 cm (inverse). If any published data is available, that will help me a lot! This is in turn an attempt to study the hot water spectrums obtained from the sun (sunspots).Following
Any speculation as to whether certain species of dinosaurs had an adaptive immune system?
A theoretical question out of curiosity. Sorry that I lump all hundreds of species under an umbrella category of "dinosaurs". Just curious to know if there was an expert opinion as to whether Dinosaurs expressed a combined innate/adaptive immune system. If so, how would one determine that? Also, which species of dinosaurs would this apply to? Just wondering.
Thanks in advance for the responses.
Good point Daniel, since those would be the closet living relatives of dinosaurs, that would be a good start.Following
How can I calculate absorption coefficient from absorbance?
I am interested to know which formula can be used to determine the absorption coefficient (alpha) from an absorbance. Absorbance is obtained from UV-Vis measurement.
it is a powder sampleFollowing
Can anybody give me detailed step by step analysis on how to carry out moisture absorption test on polymer composite?
What is the best way to prepare high pressure die cast magnesium alloys samples?
Hi, I would like to know the best ways of preparing high pressure die cast magnesium alloys samples. Right now, I am sort of using 400, 800, 1200, 2400 for grinding, then 3 micron and 1 micron DP suspension for polishing for 15 minutes, then fine polishing with colloidal silica OPU for 20 mins. Some people who use the same samples as I, polished their samples for only approximately 5 mins, then fine polish for 3mins. Could anyone recommend the best way including the optimal time for grinding, coarse and fine polishing?
Please contact Professor K.U.Kainer.
How many type of transform can be used in EEG feature extraction?
like FFT, STFT,DWT,Stockwell Transform (S-Transform) etc.
which transform is better and why?Following
What is the difference between a pcr primer and a sequencing primer?
is there any difference between the primer used for pcr and for sequencing?
Hi! RP, am not clear about your answer if PCR primer is non specific how it anchor/binds/hybridize at specific/particular position of template DNA!!! And you get the specific amplicon!!! Could you plz explain? Refer wikipedia for basic knowledge or Campbell Biology.
"Primer sequences need to be chosen to uniquely select for a region of DNA, avoiding the possibility of mishybridization to a similar sequence nearby."
Coming to the question of NPL we use the same primers for sequencing that we used for PCR but the only difference is we perform Forward and Reverse separately using single primer for sequencing reaction to get lagging and complementary seq.s later we align them. Here the principle is Sanger's di-deoxy termination method. Where as in normal PCR we use both F n R in the same reaction. I believe you aware what is PCR: Hope your doubt is clear.Following