SDS-PAGE problem? - FAQS.TIPS

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Food chemistry book ?

what is the best Food chemistry book that I can use for preparation of lectures for postgraduate students ?

03 April 2019 3,109 2 View

Change in β-Lg during heat treatments?

In the attached SDS-PAGE photo I studied the change in SDS– PAGE patterns of the pH 4.6 whey samples separated from milk heated at different temperatures for 30 min. ( 2, unheated milk, 3,...

02 March 2019 6,115 4 View

RP and normal phase HPLC ?

Whats are the differences between RP and normal phase HPLC ? and what is the best choice for analysis amino acids in foods ?

02 March 2019 2,907 9 View

Is there another mode of inheritance for favism (G6PD) beside XLR mode?

In my practice I have seen a lot of cases of favism affecting females.

04 May 2014 5,289 0 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

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

How accurate is Image J for SDS bands analysis?

I want to check my protein expression but the sample (target protein) on my SDS PAGE shows a little difference as compared to the control. I measured the band density through IMAGE J but the band...

01 July 2024 1,841 3 View

How to troubleshoot error E1 on Biorad Powerpac Basic despite using sufficient and fresh buffer?

I have used this BIorad Powerpac Basic for 4 years now, before which it was used for another 5-6 years. Generally we addressed the E1 error by compensating with fresh buffer and/or filling the...

01 July 2024 7,104 1 View

Thin DNA gel loading sample tips?

I have a 0.25mM thick gel and I am having trouble cleaning out the wells to ensure constant sample application. I usually use a thin syringe to get into the wells and rinse them out with running...

01 July 2024 2,124 2 View

Saroj Pandey

I think you have too much protein on the last two lanes. Just try again with less samples. Might help.

Try reducing voltage during electrophoresis which also helps in better separation of the proteins.

All the best

Raheleh Tooyserkani

It seems to be a result of overloading! Also as Saroj mentioned, try to decrease the voltage and also you could increase the length of gel to improve resolution.

One more point; always try to use freshly prepared buffers, if possible.

Good luck

Jasim Al-saadi

thank you all for your answers , but I face the same problem when I decreased the volume of the injected sample .any suggestions please ?