Which acrylamide:bisacrylamide ratio would be the best for tris-tricine gels?

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Is it possible to purchase cDNA of Giardia?

We would like to sublone several genes for expression, thus the genomic DNA (sales by ATCC) is not good enough for us (since few of these genes include introns. Any suggestions?

29 May 2023 8,334 0 View

Which anti-fading is suggested for microscopy?

We experienced a fast bleaching of fluorescent staining of S. cerevisiae cells with Nile red. Wondering which anti-fading is suggested for microscopy?

24 October 2017 7,352 8 View

Disulfide bond formation causes proteins to migrate more quickly in SDS-PAGE

While using TCEP we realized that Di-sulfide bond formation causes proteins to migrate more quickly in SDS-PAGE. Does anyone have an idea how to recognize the specific cysteines involved in this...

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Does anyone know what might be causing the smeared bands on my western blot?

I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...

10 August 2024 7,480 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...

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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...

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Low-yield gel extraction problem?

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PCR showing no bands - Master mix and primers didn't mix?

Hello everyone, I performed a PCR yesterday, and the results showed no bands on the gel. Of course, I probably missed some crucial steps, like adding my samples to the PCR strips themselves, for...

31 July 2024 2,406 6 View

How to check pH of a high strength hydrogel?

We are working on biopolymeric hydrogels. Our system is highly viscous and sticky, and the gel formed are high in strength. We are unable to use pH electrode and pH strip. Please suggest an easy...

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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...

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Antonina Kokisheva

standard ratio 32:1 (including for small proteins)

http://www.hixonparvo.info/SDS4Peptide.pdf

https://static1.squarespace.com/static/570deb0755598634587afa25/t/57329292859fd0590141c67e/1462932115229/SDS-PAGE+using+Tris-Tricine+system+%28%26+Gradient%29.pdf

Elah Pick

We ended up with a 12% tris tricine gels. Thank you so much Antonina Kokisheva