Ruchi Kakar

6 Questions 43 Answers 0 Followers

Questions related from Ruchi Kakar

Co-immunoprecipitation: Interacting proteins seen in Isotype Control too?

I am trying to optimize a Co-immunoprecipitation experiment where I intend to see if 'Y' and 'Z' are interacting partners of 'X'. I am able to immunoprecipitate X and the isotype control does not...

07 July 2018 6,458 8 View

Protein over expression in stable cell clones - can anyone help?

I have created stable cell clones of a protein whose gene was cloned in pcDNA3 which has G418 resistance. I started observing colonies after 15-20 days of growing them in the medium. The cells...

10 October 2013 5,556 11 View

Protein not being detected in western blot but seen in immunofluorescence and RNA confirmed by Real-Time PCR.

Rab11, a trafficking molecule does not come up as clear bands in western blotting but shows a lot of non specific bands. Same antibody when used for immunofluorescence shows intense staining. It...

02 February 2013 2,133 9 View

Cells become round and detached after changing to fresh medium in plate

I have been trying to transfect an endometrial epithelial cell line with a plasmid. But before transfection, when I change the medium of the cells and replenish with the same growth medium in...

01 January 2013 5,882 69 View

Can anybody suggest a good lysis buffer for Rab11A protein?

I know the question is very specific but this protein even being cytoplasmic, just does not come in any lysis buffer I have tried.

05 May 2012 1,295 5 View

I am facing a problem with western blotting of a trafficking molecule which is a 25 kDa GTPase located in recycling endosomes of the cell.

I load approx 10 ug protein usually for western blotting. the problem that i am facing is that the fresh lysates that are prepared in lysis buffer (Urea, tris base, CHAPS) does not give me a band...

02 February 2012 4,206 26 View