What will be the best protocol for transfer of separated high molecular weight proteins from SDS-PAGE gel to nitrocellulose membrane?

I am facing problem in the transfer of high molecular weight proteins to a nitrocellulose membrane from SDS-PAGE gel. Experimental target protein has a molecular mass of ~125kDa, and I tried...

09 October 2015 8,366 7 View

How long we can store SDS PAGE gel before Western Blotting and what will be the effects?

Is there is any possibility that my gel will not suffer any damage/alteration in blotting, if I store the gel before western blotting?

10 November 2014 9,971 9 View

Does the volume of media affect the growth of cells in animal cell culture?

As media contain all the necessary factors for the growth of cells. The excess may result in altered behavior of cells, if I use undetermined cell culture media volume, will it affect the growth...

09 October 2014 8,354 12 View

What precautions should we take to seed an equal number of cells in a 24/48/96 well culture plate?

08 September 2014 1,022 23 View

How can I avoid slimy total cell lysate extraction?

I am facing a problem during extraction of total proteins from cell lysate. I am using cell lysis (RIPA) buffer with standard composition as described in scientific literature. But every time my...

08 September 2014 1,612 6 View

How to decide dose concentration for animal cell culture experiments?

As for a particular range of treatments we observe a hyperbolic/bell curve. Which region do we select from the graph for experiments? Is there any particular criteria for dose concentration...

07 August 2013 6,446 4 View

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02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

01 March 2021 8,087 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View