Shivasankari Gomathinayagam

10 Questions 29 Answers 0 Followers

Questions related from Shivasankari Gomathinayagam

Protein-DNA complexes stuck on the wells of native PAGE during unwinding assay.

I am doing an unwinding assay for my protein with a fork substrate and my protein-DNA complex is stuck in the wells. My quench to stop the reaction has 0.5% SDS, unlabelled strand, EDTA and...

05 June 2017 3,875 10 View

What is the best way to store Rabbit serum for long term use ?

We raised our Antigen specific antibody in Rabbits and we are planning to use the serum without further purification. My western blot bands are already specific without much background, hence the...

22 December 2016 3,108 4 View

Can someone help me with Immunostaining of suspension cells?

I am doing a immunofluorescence imaging of my suspension cells (EBV lymphocytes) to look for RAD51. However for some reason I am not able to see any RAD51 in the cells. The DAPI stain works but...

22 November 2016 1,798 6 View

How to prevent clumping of EBV trasnfected lymphocytes?

I am culturing lymphocytes and they tend form visible clumps.. Does anybody know how to avoid this and if this is common ? I am new in culturing them. Thanks

31 May 2016 800 7 View

Help with imaging Western blots on an imager?

I am new to using the imager for my western blots. I am not satisfied with the images I get. I would like to know how to get good images using a imager. 1) Do you pour the substrate on the top of...

30 November 2015 9,886 10 View

Which antibody could I use against BRCA2 for Immunofluorescence?

I am looking for an Antibody to use in Immunofluorescence specific for BRCA2. I am not able to find a convincing IF image of BRCA2 by IF. Lot of scientists use the tagged version of BRCA2 in their...

16 September 2015 3,963 3 View

Why are my U2-OS cells look different i.e. long like nerve cells?

We received a vial of U2-OS cells from another lab and I put them in a 75cm2 plate to let them divide. I see that the cells are really long, I have not seen such a morphology with U2-OS cells...

13 July 2015 6,492 2 View

What is the best procedure to purify Ab from rabbit test bleed ?

I have test bleed from immunized rabbits and would like to purify my Antibody of interest from it. I was thinking of the following steps based on literature analysis : 1. Ammonium sulphate...

27 May 2015 4,414 11 View

How can I stabilise a protein-DNA complex in isolation?

I have purified my protein-Holliday junction complex by SEC, but the purified complex is not stable. The protein dissociates from the DNA. I would want the complex to be stable to perform further...

24 January 2014 6,616 2 View

Has anybody experienced protein sticking to the frits of the Superdex 200 column?

I am working on a human helicase protein and when I load them on a SEC column Superdex 200 (GE), I don't get a good recovery. In addition, when I load my protein-DNA complex into the same column...

22 January 2014 6,410 3 View