Manisha Shahu

6 Questions 8 Answers 0 Followers

Questions related from Manisha Shahu

Can a fluorescence microscopy could be use to determine a colocalization of a protein instead of confocal microscopy?

I want to determine the colocalization of a transmembrane protein along with a membrane marker, could it be possible to obtain the result with fluorescence microscopy alone or it is must to have a...

04 July 2023 2,682 5 View

Does GFP get cleaved from a fusion protein ?

Hi, I am encountering a problem with protein expression and than detecting on western blot. I have around 120kDa membrane bound protein which is now fuse to GFP(~150kDa) to help in selecting...

14 March 2023 6,115 1 View

How to improve ladder like background in western blot?

Hi, I am trying to analyze the protein expression of 2 different proteins via western blot. Since both are different isoform I am using two different antibodies for each respectively. However, in...

16 January 2023 2,810 4 View

Is it possible to perform PCR without DNA isolation?

Hi, I am trying to do a PCR from transfected HEK cell containing my protein of interest, however I am not getting any success, while the control DNA is giving the correct band with same primers...

23 November 2021 7,526 4 View

How to troubleshoot loading insolubilize transmembrane bound protein on SDS gel for western quantification?

Hi, I am working on a membrane bound protein, but due to experimental approach of checking its activity I cannot solubilize my protein and thus I do not use any detergent in my lysis buffer....

03 May 2021 9,273 2 View

At what stage do I need to do FACS cell sorting when generating stable clone with transfection?

Hi, I am trying to create a stable cell line with my protein of interest inserted in vector expressing GFP. However my protein is not directly attached to GFP expressing region. I would like to...

04 January 2021 1,144 5 View