Contact experts in SDS-PAGE to get answers
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Questions related to SDS-PAGE
I am currently working with COS-7 cells. I have noticed recently that every time I passage them, I find a lot of dead cells 2 days later. I use DMEM with 10% FBS and trypsinize them for 5 minutes...
03 February 2021 6,870 3 View
A protein dimerizes through CTD. I have expressed CTD of protein with two different tags. Now, I want to show that these CTD of protein dimerize in a pull-down assay. But my doubt is that if the...
30 January 2021 7,676 4 View
When making the SDS-PAGE running and stacking gel solution, can I save the solution without adding the TEMED and APS for future experiments? If so, how long can I save it?
29 January 2021 3,930 3 View
Hi everyone, We are doing western blot with yeast protein extracts. We are applying a wet transfer method using Tris-Gly as a transfer buffer (with 10% methanol) and running it with constant...
26 January 2021 4,326 6 View
I ran SDS-PAGE with 14% lower gel, 4 % upper gel, having brushing, pre-running empty gel for 30 min. The protein samples had been centrifuged with 14,000g for 10 min, and then the supernatants...
21 January 2021 312 3 View
I ordered a synthetic peptide (sequence = MHHHHHH) This peptide has a molecular weight of 0.97 kDa based on a web-based prediction tool and mass spectrometry. However, when I ran it on the...
21 January 2021 3,576 4 View
I am working with an Fe S cluster proteins and it seems to be in inclusion bodies, even after avoiding sonication, doing lysozyme lysis with DNAase and varying IPTG concentration. It is not a...
21 January 2021 8,970 5 View
Are there companies that do protein extraction, SDS page and Western blotting on tissue samples? I have one target protein and would like to know the concentration in each sample.
20 January 2021 890 4 View
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 10,079 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 7,204 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 3,294 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 2,046 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 7,895 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 4,542 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 7,389 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 546 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 4,724 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 2,907 3 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 3,714 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 1,948 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 4,795 5 View
03 December 2020 5,114 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 1,555 3 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 9,094 1 View