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Questions related to Proteins
Hi All, I have been doing a wet transfer for my 10 % gel in a NC membrane for a protein at 42kDa at 100V for 1h with a freshly prepared transfer buffer substituted with 20% methanol. After my...
08 August 2017 5,878 2 View
Is there a way to calculate the backbone RMSD for two different peptides that have the same number of residues but one of the side chains is different? It should be possible because the two...
06 August 2017 1,296 3 View
I am working with a pET30a+ construct and my protein is C-terminal His-tagged. My protein is forming inclusion bodies and I need to get a protocol for isolation of a pure protein from this. An...
01 August 2017 5,041 3 View
Hello, I'm trying to purify a 47 kD fusion protein by loading the supernatant directly on the column but I'm having issues with multiple species in my elutions. The lanes are: ladder,...
14 July 2017 5,334 6 View
Granted, there are inclusion bodies that contain huge concentrations of small bioactive proteins that can be isolated without loss of bioactivity. But trying to isolate large proteins and...
07 July 2017 8,494 20 View
Hi ! I expressed a membrane protein in Lactococcus lactis and usually the lysis is carried out with a french press. But I have a small volume (1 mL in epp tube), the only possibility for me is to...
07 July 2017 9,813 2 View
In experiment, We generally use formaldehyde to make DNA and proteins form complexes, so, I have a question that what is the real role of formaldehyde(FA) on the effect of crosslink between...
07 July 2017 4,592 2 View
07 July 2017 7,067 7 View
Hi all See the SDS PAGE gel below stained with an enhanced Coomassie dye, Imperial Stain. I centrifuged a vial of Collagen IV. 30ul of the supernatant was loaded on the middle lane. A portion of a...
07 July 2017 6,435 8 View
Hello, I've observed a curious phenomenon that I need help with understanding. I have a hydrogel sample that is ~3% protein, ~3% polysaccharide (unknown size), in saltwater. When this sample air...
07 July 2017 2,731 3 View
The crosslink of a purified protein to NHS-activated magnetic beads works absolutely fine. But now I want to crosslink human skin tissue lysate to the beads and this does not work at all. The...
07 July 2017 3,230 3 View
06 July 2017 7,814 3 View
01 July 2017 3,371 3 View
Hello, Is it possible to separate a complex fluid in a vesicular part (containing vesicles / exosomes) and a non - vesicular part, containing other proteins? Many thanks!
29 June 2017 5,598 3 View
After DSF guided refolding of protein, results for denatured and refolded samples are coming out like this. is it ok if the denatured sample give peak and Tm like this? as i have used mild...
24 June 2017 1,863 3 View
As part of an assay, I need to re-suspend acetone precipitated protein samples from whole heart tissue in a 1 mL RIPA buffer that has 1% SDS. I'm having great difficulty breaking up the tissue, so...
22 June 2017 5,785 4 View
For the expression of subtle Nus tag fusion protein I have optimized sonication for 5ml culture, 50 percent amplitude, 1pulse off 1pulse ON for 30 second. With repeated cycles with the gap of...
21 June 2017 1,425 9 View
I have the list of proteins which are either inhibited or induced by action of certain small molecule. I would like to know the downstream effects of this action so that I can hypothesise a mode...
21 June 2017 1,298 2 View
I don't really understand the concept of Biorxiv. What is it? Why do people use it? Are there any benefits or downsides of preprinting?
19 June 2017 6,563 25 View
I need bacterial and virus antigen from bacteria and virus infected leaves, respectively. Can i use any protein extraction method for this purpose irrespective of pathogen type? I am confused that...
09 June 2017 8,893 4 View
tried rosetta but didnt get expression.
09 June 2017 8,664 2 View
I have a Pichia expressed and secreted protein, which i need isolate. Anyone knows how to purify the protein from the media in a large volume setting?
08 June 2017 8,194 2 View
I am planning to extract total protein from tumor tissue and to coat it as antigen for Indirect ELISA application. Please let me know what buffer I should use to dissolve the protein so that it...
08 June 2017 6,998 5 View
I need to study the effect of the protein Y mutation on the levels of a protein X. So I over-expressed the protein X under 5 conditions: (1) not over-expression of protein Y (2) with...
08 June 2017 5,343 4 View