Does anyone know the reason I can find very faint bands in SDS Page whereas no bands in a native page?

02 February 2014 1 5K Report

I am trying to run pepsin on sds-page/native page.

What amount of protein you are loading in well?? I think you should increase the quantity of protein about 50 microgram. Also standardize your staining method.

Badges
Science method

More Aniruddha Bhati's questions See All

Is there any significant role of silica in using a silica magnetite nanoparticle for extraction of bio-molecule like DNA?

Explain the mechanism please?

01 February 2014 9,693 3 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

Why is my ASO degraded quickly on agarose gel?

I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...

06 August 2024 3,130 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

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...

01 August 2024 4,673 4 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

Why my gel electrophoresis has smearing?

Greetings. I’m currently running a nested pcr for giardia. My mastermix comprised of 3mM Mgcl2, 5unit of taq polymerase, 0.2uM of forward and reverse primers each respectively and 0.2mM of dNtp...

22 July 2024 9,761 5 View

How should we increase the quality of RNA extraction?

I’m having difficulty achieving high RNA integrity in my samples. Although the 260/280 and 260/230 ratios are satisfactory after RNA extraction, the RNA samples show signs of degradation when...

22 July 2024 155 4 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

Why my gel electrophoresis have shadow bands? Please see the attached picture for the gel electrophoresis ?

Sometimes I see the shadow like bands and its not true band. I want to know that what's the reason for it. I am using 2% gel for running genotyping samples I have uploaded the gel picture in both...

19 July 2024 148 6 View