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Questions related to SDS-PAGE
I have tried SDS_PAGE and native PAGE in my laboratory with a locally purchased gel casting system from MEDOX, India. I never got a clear distinct ladder even for the molecular weight marker. i...
15 April 2020 2,904 8 View
I am purifying a recombinant protein (MW 37 kD) using IMAC. I used the Bradford colorimetric assay (by adding a drop of the solution from the column to Coomassie blue G-250) on the wash and...
08 April 2020 3,264 9 View
We are following the protocol of mammalian tissue and Mouse/Rat to extract DNA from fish (page 28, 23 and 24): https://assets.thermofisher.com/TFS-Assets/LSG/manuals/purelink_genomic_man.pdf, but...
02 April 2020 413 0 View
I know both can be used, but why is SDS-PAGE "preferred"? Does it have to do with cross-reactivity?
30 March 2020 6,153 5 View
Dear researchers, How to make elution buffer for SDS-PAGE? There is a lot of protocol to make elution buffer, but they didn't show the amount of each ingredient needed. thank you.
26 March 2020 8,473 4 View
I have purified enzymes cellulase and beta-glucosidase and performed SDS-PAGE. Therefore, I have characterized with metal ions at 2mM, 5mM and 10mM but at 5mM concentration lower activity than at...
26 March 2020 8,183 3 View
Dear researchers, May I know do anyone can suggest the protocol of protein elution from SDS PAGE ? Any simple and inexpensive methods? Thank you.
25 March 2020 2,196 13 View
Do I look at the most prominent bands on the figure? or do I look at bands at level 53kDa, even though they are not the most prominent in the image? Would i also mention that multiple bands...
24 March 2020 2,547 3 View
I'm working with some short protein with size around 20 kDa. I'm expressing the protein in E. coli BL21(DE3) system at 25 degree, 150 rpm in YT media. While I'm checking the proteins in cell...
18 March 2020 2,458 3 View
Hello, I use homobifunctional crosslinkers to form protein gel. I am currently using 4-nitrophenyl carbonate leaving groups for protein crosslinking. I dissolve cross-linker in DMSO and slow add...
17 March 2020 9,501 3 View
The flour I am testing is quite coarse and the samples are already quite hard to dilute. I tried different dilutions, but after 5 min heating they still become partly solid (like soft ball of...
16 March 2020 5,526 3 View
I try to resolve collagen on SDS-PAGE. My goal is to resolve the collagen extracted from skin scaffolds. I use ThermoFisher Bolt mini gels. I can see the protein ladder migrating and also I can...
16 March 2020 10,013 8 View
Hi:) I'm doing His tagged peptide expression and purification from E.coli. I expressed the peptide and checked it with WB (with Anti-His Ab). For purification, I did IMAC and got an eluate. But it...
13 March 2020 6,487 13 View
I am wroking on microbial enzyme and after its extraction in crude form, I had performed the SDS PAGE. But I was not able to get any visible band of protein instead of repeating the whole...
05 March 2020 2,053 4 View
Hi, I recently purchased a MidiPrep Kit, and the protocol said at DNA precipitation stage, I need centrifuge the sample 12,000 at 4 degree C. The problem is my lab does not have this...
28 February 2020 9,644 1 View
Hi there, I'm trying to purify some potentially interesting native secreted protein for a bacteria. I can see some good strong peak on the UV trace following IEX chromatrography using an AKTA...
26 February 2020 226 3 View
I have cloned Hemagglutinin (HA1 only) with 6xHis on its C-terminus into pet23C and expressed in E.colli (Bl21(DE3)) and then purified on Ni-NTA column, but my protein after purification had...
25 February 2020 1,914 5 View
I am doing an SDS PAGE with LAD2 cells lysate but the total protein obtained was 0.4ug/ul when 200ul of RIPA was used to lysed 5 million cells. Sample was lysed for 45 minutes on ice. Kindly help.
25 February 2020 5,870 3 View
Dear researchers, I would like to carry out dialysis on my plant extract before carrying out SDS-PAGE. Beside 0.2M sodium acetate buffer (pH 5) can as dialysis buffer, may I know which dialysis...
19 February 2020 1,874 2 View
After successful purification of my protein of interest by affinity chromatography (first picture, right side), yielding 50ml of 0.4 mg/ml (estimated by nanodrop), I went on to perform...
14 February 2020 3,897 4 View
Dear, Whenever I run an SDS-PAGE, I get an unwanted vertical line appearing at the bottom of the gel. Does anyone have any idea why this happens? Any comments or suggestions would be highly...
11 February 2020 8,599 1 View
Dear all I extracted LPS from E. coli outer membrane and run them on vertical gel. And it showed a ladder-like pattern of bands, I wonder what these bands represent respectively? If each part of...
03 February 2020 8,171 2 View
02 February 2020 5,636 5 View
02 February 2020 9,602 4 View