SDS-PAGE

Contact experts in SDS-PAGE to get answers

1,312 views 2,322 posts

Questions related to SDS-PAGE

Any suggestion on SDS PAGE ?

Dear researchers, I have a problem on SDS-PAGE. My protein concentration of plant protein extract is about 0.7032 µg/µL. After that, I mix 28µL of sample with 7µL of 4x sample buffer to load...

28 September 2020 8,848 10 View

Why am I getting multiple bands in western blot (in Isolating adhesion complexes) ?

Hello everyone, I am isolating adhesion complexes and wanted to confirm the presence of a specific protein in the isolated complex. When I performed western bott, I got a band at the right size...

20 September 2020 679 7 View

How to induce with L-arabinose Chaperone Competent Cell BL21 (Takara)?

I am using Chaperone Competent Cell BL21 from Takara, pGro7/BL21 , but it is not clear on the protocol how and when I should induce the expression of the chaperones with L-arabinose. First time I...

15 September 2020 8,652 3 View

Why appear two bands of protein in sds-page after the purification?

Hi guys, I am purifying a protein by Ni-NTA column, but I am getting two bands in SDS-PAGE. What do you know to fix this problem? Please.

13 September 2020 1,862 13 View

Why SDS PAGE bands are coming like this?

Hello everyone, Hope all are safe. I have done SDS-PAGE for my desired protein from a microbial source and immunoglobulin. In this regard, I have loaded 100 microgram protein in each well, but...

09 September 2020 774 15 View

Membrane protein missing from SDS-PAGE gel?

I am trying to run an SDS-PAGE after his-tag purification of a sample. The protein of interest is an oligomerising membrane protein with an YFP tag attached. When the fractions of the sample from...

08 September 2020 3,394 6 View

What is the best program for detection of proteins analysis on the gel in the SDS-PAGE?

SDS-PAGE is used to separate the particles in the mixture, under the influence of the electric field applied to it.

07 September 2020 5,418 3 View

Desired membrane protein band isn't aligned with marker in SDS-PAGE gel. What could be the possible reason?

I am having problems regarding to protein separation via SDS-PAGE. As a newbie in this field, I have tried to optimize the conditions for protein induction. My protein size is in 20-25KDa range...

04 September 2020 8,349 11 View

Feline immunodeficiency virus (FIV) radioimmunoprecipitation assay (RIPA) protocol?

Can someone share an FIV RIPA protocol? I have FIV infected/control cat sera that I want to check for antibodies to FIV proteins. Looking for a cell transfection with...

01 September 2020 8,569 1 View

Why is my protein a smear when I detect with CoIP lysis buffer plus LDS?

Hi I am trying to express my protein R in tobacco leaves and do a CoIP, but I can't detect the protein R in CoIP input. I took the same tissue and added 8M Urea plus LDS protein buffer, protein R...

01 September 2020 9,657 10 View

Why does my gradient SDS-Page gel turn white when put in Coomassie blue staining solution?

Can anyone explain why a gradient gel (bought from BioRad) would turn white when put in Coomassie staining solution? I have never had this problem before with gels. I have always added the...

27 August 2020 2,344 6 View

Can an isolated protein domain give a single band in SDS-PAGE equivalent to its dimeric mol. wt. ?

The protein I'm working with forms a homodimer and is a 2-domain protein. I have cloned and purified one of its domains. However, when I run the domain on SDS-PAGE, I get a single band equivalent...

20 August 2020 8,031 14 View

TCE Stock solution concentration for SDS-PAGE?

Greetings, I'm trying to standarize TCE free staining method to visualize protein bands over SDS-PAGE, it's never been used in our lab. I've seen many published papers using this method, and the...

18 August 2020 6,789 3 View

What's the main difference between #Tris-glycine #SDS-PAGE and #Bis-Tris #SDS-PAGE. How pH and ion strenght affect SDS-PAGE?

sometimes, there is some smear on the SDS-PAGE, this can be eliminated with a different running buffer or a different PAGE, or a different way of sample loading buffer. how these affect the SDS-PAGE?

10 August 2020 6,281 3 View

Must I remove Triton X-100 and/or NaCl before before Bradford assay and/or SDS-PAGE (with BME)?

Hello! I'm purifying a protein and have several questions about what is appropriate to include in the extract solvent at different stages of the purification and characterization phases. (1)...

06 August 2020 4,747 7 View

What is the problem with my blot?

Why are my cell lysate sample not running properly. This is ponsue staining just after transfer. I did coomasee stained of my SDS PAGE gel and have seen similar effect. Acn anyone suggest what is...

30 July 2020 4,864 9 View

Anti-His Antibody binding everything on a western?

Hello all, I've been expressing some His-tagged proteins in E.coli that appears to be going just fine based on how the SDS-PAGE gels look, so I wanted to confirm with a Western. I transferred my...

23 July 2020 5,357 5 View

Why there is a 11 kDa band showing in SDS-PAGE of my IgG1 digestion with Papain?

Hi guys, I am using the papain digestion system to abtain a Fab. Here is my recent result, I got thin bands around 55, 45; thick bands around 30, 25, which may indicate incomplete digestion, but...

28 June 2020 3,985 9 View

Can anyone tell me if why i am getting my recombinant protein both in supernatant and pellet after lysis ?

hello all I am expressing a His6x recombinant protein and after lysis in lysis buffer i sonicate the cell suspension (40% amplitude 30 sec ON 30 sec OFF , 5 cycles). when i run SDS PAGE of the...

19 June 2020 5,601 12 View

Why can't I see 25kDa lower bands in my 15% SDS-Page gels?

I have done multiple SDS-PAGE but since few weeks I am getting a blue smear that does not allow to see bands lower than 30 - 25 kDa. My gels are 15% Polyacrylamide. I have change all...

17 June 2020 8,785 6 View

Could anyone give me any suggestions for loading gliadin/ Glutenin in SDS-PAGE well ?

I used 5 ug gliadin and loading dye ( total 10-15 ul) but it’s come out . Then modified it to 2.5 ug gliadin again faced same problem . Before loading I followed all steps ( extractions and...

08 June 2020 3,958 11 View

Someone has a suggestion to improve the purification of MMLV-RT protein?

I am working with a MMLV-RT (Moloney murine leukemia virus- Transcriptase Reverse). I am expressing in E. coli pTGroE and purifying in a Chelating Sepharose loaded with Ni2+. I am eluting the...

02 June 2020 4,205 4 View

What is the reason for smears in cell membrane SDS-PAGE?

Hello there; I'm trying to optimize SDS-PAGE/ Western blot condition for Eukaryotic cell membrane. After tissue separation, resuspending in RIPA buffer and sonication, I centrifuge samples at...

05 May 2020 3,180 3 View

What is the best brand of protein molecular weight marker for SDS-PAGE?

Hello, I use from Biorad Precision Plus Protein dual-color standard (MW marker), but the molecular weight that is estimated with this marker isn't usually precise. to confirm, I used from USP...

20 April 2020 3,228 5 View

Badges
Science method