Ashish Kumar

10 Questions 10 Answers 0 Followers

Questions related from Ashish Kumar

Fluorophore proteins (eg GFP, YFP, RFP) expressed in bacterial expression system are active?

Hi everyone, I have to express different mammalian proteins tagged with different fluorophores (such as GFP, YFP and others) under bacterial expression system and to use these purified proteins...

04 April 2019 1,555 4 View

FRAP Vs iFRAP ?

Hello everyone, Please suggest me if FRAP and iFRAP results complements each other or not, because FRAP deals about the recovery kinetics and iFRAP for dissociation. Should the mobile fractions,...

10 October 2015 1,915 0 View

Anyone familiar with nucleolar localization of proteins?

How does the nucleolar proteins/ribosomal proteins find its way to the nucleolus. Like nuclear proteins have NLSs and NESs which are responsible for their import and export in the nucleus...

04 April 2015 5,205 2 View

Is anyone familiar with BrdU / BrUTP incorporation?

I have a query on BrdU/ BrUTP incorporation in culture cells. Why is transfection/microenjection required for BrUTP incorporation. Whereas, We add BrdU directly into the culture medium to pulse...

10 October 2014 5,592 7 View

How can I use 2 different antibodies (for two different antigens) from same origin in IFA?

Can you please suggest me on how to use 2 different antibodies (for two different antigens) but from same origin in IFA. Both the antibodies are from mice. I tried the staining sequentially for...

05 May 2014 4,511 13 View

Can someone please tell me what protocol I should use to re-isolate RNA after DNAse treatment?

I use Trizol method to isolate RNA from frozen PBMCs. Sometimes, I get 260/280 ratio of ~1.8 at which I get slight amplification in my no-RT control, indicative of genomic DNA contamination. I...

04 April 2014 9,130 15 View

For recombinant protein expression in E.coli is Shine-Dalgarno (RBS) sequence mandatory?

I am cloning a protein in a bacterial expression system, but I cannot use RBS given in the vector, so I need to add it to my forward primer. Is it absolutely necessary? What is the optimum length...

12 December 2013 4,450 12 View

Nuclear protein isolation?

I am isolating proteins from crude nucleus using high salt concentration( 300-500 mM NaCl). Basically, I have to get nuclear proteins which will be used for CO-IP. In protocols I tried- a) 0.2mM...

11 November 2013 5,521 5 View

Cell cycle..

hi everyone...i have to see the effect of certain chemical compound on the cell cycle..basically i have to see their effect on the check points,whether it is causing cell cycle arrest at any...

09 September 2011 8,047 1 View

Methods to study a competitive protein-protein Interactions.

What methods can be used to study a competitive interactions between two different proteins (eg. A & B) for a common binding motif on protein (C).

01 January 1970 9,648 4 View