Contact experts in SDS-PAGE to get answers
1,768 views 2,488 posts
Questions related to SDS-PAGE
It is well-established that size effects due to large strain gradient effects play an important role at this scale. How can we avoid this effects? Ashish
02 May 2021 7,193 0 View
Hey everyone, I have purified GST-tagged protein in elution buffer (200 mM Tris-HCl pH 8, 20 mM NaCl, 40 mM reduced glutathione, 10 % Glycerol) I read that the glycerol can be problematic for BCA...
29 April 2021 3,350 8 View
Hello all, so I'm trying to purify a protein complex with multiple subunits for the very first time, and after eluted with different concentration of imidazole there are multiple bands on my...
27 April 2021 6,656 5 View
What happen that caused these white spots/build up? Just some background if needed, I cast my own gels and I ran two 15% SDS Page gels today. One of them came out perfect while my second gel...
24 April 2021 8,112 3 View
I will try to be as thorough as possible. So it's been over a year or so and I went back to pouring various % SDS Page Gels and I keep running into problems I didn't have before as seen below. The...
20 April 2021 3,187 3 View
Polyacrylamide gel that is a component of SDS PAG, my concern is to figure out the concentration of polyacrylamide gel that can be used for SDS PAGE.
20 April 2021 4,210 3 View
I used to dissolve my sample of collagen in different solutions such as 1M NaOH, 0,1M acetic acid, distilled water befor mixed it with laemmli buffer. And also I tried a different gel...
18 April 2021 7,051 2 View
Target bacterial operon is reported to be co-transcribed and co-translated in wild type host. However, protein expression analysis (SDS PAGE) indicates translation of only first gene in frame...
16 April 2021 3,908 3 View
When I was doing a SDS-PAGE upto 25kD the bands of the protein ladder got resolved properly , below the 25kD the 20, 15 and 10kD bands did not resolved and the tracking dye of the samples was...
13 April 2021 8,833 6 View
Does anyone know if there is any condition or modification of protein that will prevent the protein band to be shown on SDS-PAGE gel after silver staining? I recently produced a human protein...
09 April 2021 6,867 3 View
Hello I use the Bradford method to measure the amount of protein expressed by E.coli BL.21. My problem is that despite the R² = 0.9814, the average absorption read at 6 and 21 hours after adding...
09 April 2021 2,104 4 View
Hi all, I've been working with HL-60 cells for ~2 years in our lab. I've been able to successfully differentiate them to a neutrophil-like phenotype using DMSO (1.3% final concentration, 5 days...
06 April 2021 1,840 3 View
Both hydrophile and hydrophobic can enhance the boiling performance in some researches, is there relationship between the wetbility of surface and the boiling performance? How to explain?
31 March 2021 5,178 3 View
In order to produce a chimeric antibody, I transformed HEK293T cells with two plasmids: one for heavy chain and the other for light chain IgG. The supernatant was purified in a Protein G-Sepharose...
29 March 2021 4,693 8 View
I'm carrying out B-cell isolation from mouse tissue by cell sorting using 2x biotinylated probes. One is a wild-type protein and the other has a single point mutation to reduce binding of B-cells...
26 March 2021 4,441 3 View
Does anyone have a suggestion on how to identify/see peptides on SDS-Page gels? I'm using the correct gel, but Coomassie just isn't strong enough for me to see the band.
21 March 2021 6,447 5 View
Dear researchers, My research is to determine which peptides pose antiviral activity. I used Acrylamide: Bis-Acrylamide Ratio 13:1 as this is ideal for Peptide separation and Color Marker...
19 March 2021 4,096 4 View
SDS Gel doesn't solidifies even if: - APS was made freshly - TEMED is not expired - Increased Acrylamide concentration Kindly help me if anyone have encountered similar kind of problem.
18 March 2021 8,053 8 View
I have an excel file of differentially expressed genes that includes the logFC and the logCPM. These are a prior student's data so I did not create them and I do not need all of the genes, I only...
17 March 2021 1,700 2 View
Have been finding these weird-looking objects in my culture dishes lately. There are several strange things here, but I am most concerned with the spore-like structure near the center of the field...
16 March 2021 9,138 11 View
Hi My protein of interest is only expressed in E. Coli insoluble fraction. So, I tried to extract them adding 6 M guanidine to the insoluble pellet. By rotating the mixture of pellet and solution...
12 March 2021 1,556 3 View
I have obtained these results from an SDS-PAGE. I am not sure how to analyse these results. What is the analysis that can be made and how do I determine the purity of the protein?
08 March 2021 8,825 11 View
I am trying to Induce a construct (1.3kb) from Drosophila in the pGEX-4T1 vector in BL21 cells with IPTG. IPTG induction was done in 3hr grown culture and induce for 3 to 4 hr at 37*C. But there...
06 March 2021 2,532 4 View
Hello everyone. I am fairly new in protein chemistry field. I am trying to perform a protein induction hoping to purify my protein of interest using FPLC method. However, I am having trouble on...
04 March 2021 8,624 3 View