Contact experts in SDS-PAGE to get answers
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Questions related to SDS-PAGE
Hello, I would like to detect the protein bands in the SDS-PAGE gel by Acid blue 92. I checked many articles but still could not find an appropriate protocol. Does anybody have experience with...
17 January 2021 10,088 3 View
Hello, I have cloned my gene of interest in a bacterial expression system i.e. BL 21 Gold (DE3) and checked for protein production and solubility. Good amount of protein was observed on SDS-PAGE...
16 January 2021 7,227 5 View
When working with SDS-PAGEs I used a 26 well instead of a 18 well 12% bis-tris for AAV work. Would this prevent the protein from moving through the gel as expected. I usually use an 18 well...
14 January 2021 3,308 2 View
What's the criteria of comolete lysis of E.coli when breaking them using ultrasonic? What phenomena can help to determine if the cell are broken completely? From smell, transparence of the lysis...
08 January 2021 2,076 3 View
Samples (constituted by Laemmli buffer 4X, total protein extract and water) were loaded after denaturation at 70 ° for 10 min. Bromophenol blue reformed an unique band only towards the end of the gel
07 January 2021 7,921 4 View
Is there anyone who has experience on SDS-PAGE with a small peptide (MW =~ 1- 2 kDa)? Is anyone able to offer advice on techniques?
04 January 2021 4,566 3 View
I have been expressing a protein with a tag which is then removed with a protease and repurified. Both purification steps use a Ni-NTA column. I run the samples on SDS-PAGE (15%, self-poured)....
28 December 2020 7,400 1 View
Greetings.. I am using Akta Pure system for my protein purification. I have optimized buffer system for protein elution. However, this time I got broad peak while eluting sample. For...
25 December 2020 575 7 View
I’m trying to pull down and purify GST tagged protein (target sequence is inserted in pGEX4T1 vector) from BL21 Cell lyset. Target Protein expresses in BL21 cells in good amount, confirmed by SDS...
19 December 2020 4,758 6 View
Hi! I study the effect of formulation excipients on protein degradation pathways and was looking disulfide-mediated aggregation by comparing reduced/non-reduced SDS page samples. I am using TCEP...
18 December 2020 2,934 3 View
Thrips ( Scirtothrips dorsalis) are devastating pests among the most significant agricultural pest in Asia. Please let us know how we can control thrips in a sustainable way.
08 December 2020 367 0 View
To determine protein concentration of unknown samples (whole cells lysate) coming through the standard curve that has to be plotted before, I know that that can be obtained after multiplying the...
07 December 2020 3,732 3 View
Hi all, I want to characterize a product-related impurity band appearing in SDS-PAGE analysis. I excised the band and incubated it in elution buffer containing 0.1% SDS. Now I want to analyze the...
05 December 2020 1,968 3 View
I'm using a standard SDS PAGE protocol, but since a month or so, some mysterious issues started to cause us trouble as seen in the first two pics (gel1 and gel2). The third picture is from a...
03 December 2020 4,818 5 View
03 December 2020 5,124 6 View
This is our first time doing SDS-PAGE, and therefore we would like to hear from more experienced people if we interpret the results right. Attached is a picture of one of our gels with our...
02 December 2020 1,581 3 View
In screening detergents that solubilize my protein of interest from IM preps in E. coli (after overexpression from a pET vector), I found that Na-deoxycholate and lauryl sarcosinate are the most...
02 December 2020 5,890 6 View
The most likely method I'm looking at is the TCA precipitation method, however the majority of the papers on this that then use Kjeldahl are old and use filter paper as opposed to centrifugation...
30 November 2020 9,105 1 View
I need to transfer two very basic proteins (pI of 11.4 and 11.04) and would like to know what transfer buffer is most suitable for these proteins when run on a denaturing gel. CAPS buffer has the...
27 November 2020 7,727 3 View
I am having trouble replicating The reported binding energy of a structure. when using quantum espresso I get a value roughly .5x that of the reported energy. Is there anything I need to keep in...
26 November 2020 7,145 2 View
I am doing an experiment on proteins. I precipitated protein using TCA- acetone method and want to check purity on SDS-PAGE. I used 25-30 microgram protein for one well, after running SDS- PAGE,...
25 November 2020 8,999 3 View
I want to purify a fragment of my interest gene. For that I have cloned my fragment into pET28a vector with his tag at its N-terminal, and transformed into both rosetta and BL21 strains. On...
23 November 2020 7,534 13 View
I purified an MBP fusion from yeast using a simple Tris-based elution buffer and denatured in SDS-PAGE buffer. Another protein co-purified with it. I noticed that heating at 95° caused this...
23 November 2020 756 3 View
Recently I have been involved in this problem: After expressed by adding IPTG, lysed with B-PER, and resolved with Buffer B (8M urea), incubate with Ni-NTA beads, everything looks fine. When I...
22 November 2020 7,602 3 View