Hi can anybody please tell me how can we elute the intact protein from SDS-Page gel...?? is there any standard protocol available ??

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Can anyone suggest me any software or bioinformatic tool to identify whether unknown or hypothetical protein have protease domain in it or not ?

I have tried ProtIdent but could not rely on it completely

01 February 2019 7,521 3 View

How can we resolve two proteins having size difference of 1 kda by electrophoresis? which type of gel should I use ??

Basically I have two proteins of 57 kda mol. weight. One is wild type and another is cleaved one . The size difference of two protein is 1 kda. I have tried different percentage of SDS-PAGE gel...

01 February 2017 7,833 17 View

Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

How to preform densitometry on SDS-page bands?

I ran a SDS-page of a bacterial lysate and I want to quantify protein concentration in a specific band. I was thinking of using a standards ladder or make some standards are different...

05 August 2024 9,805 3 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

What is the best blank for nanodrop if I want to read a recombinant protein concentration?

Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC

01 August 2024 967 3 View

Following click reaction in cell lysates, protein is immobile and remains at the top of the gel in SDS-PAGE?

I am using CuBr/THPTA for a click reaction in total cell lysates. I am facing issues with my protein sample in non-reducing SDS-PAGE where it's not migrating properly and most of it remains at the...

29 July 2024 950 4 View

Why my IgG antibody looks fragmented on non-reducing SDS-PAGE?

I am performing IgG purification and I have to show my results on SDS-PAGE. I use 10% tris glycine gel and prepare the samples under non reducing conditions. I am new to antibodies and therefore...

23 July 2024 6,664 6 View

What is the cause of this aggregation just bellow 70 kDa mark?

Hello, I was running a 12% SDS Page electrophoresis on few granulosa cell samples and got this result after the ponceau staining. The total protein lysate seem to aggregate at 70 kDa ladder mark...

21 July 2024 5,128 4 View

Less substrate signals towards the last lanes of the Native PAGE?

I have been running native page for FAM DNA substrate ( fluorescence samples) for protein DNA binding reaction. Binding is there but towards the end of the lane , I am loosing signals...

17 July 2024 6,213 4 View

Anybody can assist me in Protein and magnetic beads seperation?

Hi, I hope you are doing well, I am working to remove DNA from protein sample by using silica hydroxode magnetic beads. I used different binding buffers but I din't get my protein band in initial...

02 July 2024 5,502 4 View

Is the wash in my IP the problem or something else?

The image shows 2 western blots. The first one is of protein A, and the second is for the flag tag that the protein is bound to. The lanes are as follows Ladder, Beads, SAB(unbound), and elution....

01 July 2024 3,183 0 View

Satish Pandey

Yes Sigma provide elution kit for protein isolation from the PAGE..

Hasan Mohammad

https://www.researchgate.net/post/What_is_the_best_way_to_extract_whole_proteins_from_SDS-page_for_mass_spect_analysis

may be you want to look at this thread, if your application is similar

Dominique Liger

Hi there,

This paper should be helpful:

http://www.ncbi.nlm.nih.gov/pubmed/22585504

Anju Bala

Thanks all for your replies.