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Questions related to SDS-PAGE
I did sds page to determine the difference in protein expression between 3 types of vaccine , but i don't have scanner or densitometer available. . so i wonder if there is a software to analyze...
03 March 2021 6,652 3 View
Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...
02 March 2021 3,077 3 View
Greetings, I want to make SDS PAGE for my sample (Tick salivery gland extract), I tried concentrations 10%, 12%. I also tried high molecular weight ladder. But the protein stopped at the top and...
26 February 2021 7,977 3 View
I have to quantify a mixture of proteins. But I can not quantify its concentration by any protein detection methods as the values fluctuate. Can i quantify the protein concentration by sds page?
23 February 2021 462 2 View
I have performed SDS-PAGE on a cell lysate (25ug) prepared with 6X LB and fresh BME followed by boiling for 10 min at 95°C, but still I see signal for different epitopes (all should be around...
17 February 2021 3,079 9 View
I ran a SDS-PAGE of my sample ovalbumin in histidine buffer in 2x Laemmli buffer without heating and it got nice visible bands. However, when I heat the sample at 77C in 2x laemmli buffer for 5...
16 February 2021 3,310 3 View
Hi everyone, I’m wondering if you could help I’ve ran a Tricine SDS-PAGE to help visualise a 2kDa glycopeptide produced by bacteria and there are several issues. The gel is a MINI-PROTEAN...
16 February 2021 3,604 3 View
I am using 29:1 Acrylamide/Bis-acrylamide solution to prepare 12 % resolving gel and 4% stacking gel. The pH of the resolving and stacking gel buffers are pH 8.8 and pH 6.8 respectively. after...
16 February 2021 1,259 1 View
I conducted an SDS PAGE electrophoresis on wheat wild varieties to separate their glutenins and measure the genetic diversity of my sample. I only ran one electrophoresis. Can a genetic drift...
15 February 2021 4,192 1 View
Hello everybody, I have a problem with SDS-PAGE and I do not know why my samples stop at the top of the resolving gel and they do not run through it. The percentage of gel is 15% and the size of...
15 February 2021 9,360 6 View
Hi everyone, I am a first year PhD student working on amyloidosis. I would like some tips and tricks to crack efficiently my current nightmare, i.e. the western blotting of the Amyloid beta and...
12 February 2021 2,794 3 View
My area of research is synthesis of metal nanoparticles and their applications focusing on protein extraction from complex biological mixtures using SDS page gel electrophoresis technique. Please...
10 February 2021 3,079 3 View
I would like to quantify a recombinant protein (with native signal sequence) secreted by Pichia pastoris using the SDS PAGE. Since my standard protein (plant origin) is not pure, I would like to...
08 February 2021 4,070 9 View
Hi all, I have got a protein, expressed in P. pastoris vector, more than 97% (SDS-page) pure in the culture medium after cells separation. Despite the protein expressed has a his-tag, Nichel...
05 February 2021 5,317 6 View
We had several times the problem after the transfer: you can detect the yellow 10 kDa Band, the blue 20 kDa Band, the pink 25 kDa Band but after only the pink 75 kDa Band. 15, 37 and 50 kDa are...
03 February 2021 5,078 3 View
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 6,881 3 View
Hi, I am growing 30 mL of FreeStyle CHO-S Cells in a 100 mL spinner flask (or spinner vessel). The protocol says to grow cells in 125 mL Erlenmeyer culture flasks at 125 rpm. I would like to know...
31 January 2021 7,530 0 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 7,712 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 3,948 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 4,352 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 347 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 3,605 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 8,978 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 912 4 View