Sumit Mukherjee

10 Questions 9 Answers 0 Followers

Questions related from Sumit Mukherjee

Why c terminally tagged 6X-His is not getting detected in Western blot?

Hi, I have encountered a weird problem. I have been trying to express a protein fragment from bacteria. This fragment has an N-terminal HA and a C-terminal 6X His tag. Following expression, I...

23 November 2020 2,669 4 View

Why no expression is seen in western blot for a C terminally tagged GPI anchored protein?

Hi,

14 September 2020 9,807 2 View

What control is needed for Acyl biotin exchange assay?

Hi, I am planning to do Acyl Biotin exchange assay on red blood cell ghosts to detect the palmitoylome of the RBCs. Also, I want to treat the RBC ghosts with 2-BP and palimotoylation inhibitor and...

03 July 2020 1,865 0 View

How could a heterologous expression affect the conformation of a non-cytosolic protein?

Hi, I am working with a protein that is typically secreted from the parasitic cells that I study. I am trying to express and purify it from HEK cells. Despite of using the mammalian signal...

02 April 2019 7,286 5 View

How to create hyper/hypo osmotic conditions?

Hi, I have a very general question. Since PBS is a isotonic solution, I was wondering if 2X or 0.5X PBS could be considered as hyper or hypo osmotic conditions respectively? I know that typically...

23 September 2017 7,395 3 View

How is superoxide production related to mitochondrial membrane potential?

Hi, I was wondering if any one could explain me the relationship between mitochondrial membrane potential and superoxide production? In order to maintain its functionality, mitochondria need to...

15 September 2017 8,092 3 View

Could high mitochondrial membrane potential block ROS export to cytoplasm?

Hi, I am studying ROS accumulation in the mitochondria of Leishmania parasites, that are defective in sterol biosynthesis. Staining with MitoSox shows that the mutants accumulate very high level...

19 February 2017 6,216 1 View

Is there a way to knock down gene expression in vivo?

Hi, I would like to carry out an in vivo mice infection experiment with Leishmania parasites where I want to knock down my gene of interest in once the parasite is in the mice. Initially I would...

17 November 2015 7,103 7 View

Does any one have experience with Clontech's inducible shRNA vector systems?

Hello, I have been trying to knock down a particular protein in Jurkat cells using the Clontech's pSingle shRNA vector system. This is a doxycyclin inducible system. Initially I followed...

16 September 2014 6,779 5 View

How could I transfect Jurkats with plasmid using any home made transfection reagent?

Hi, I would like to transfect Jurkat T cells with some plasmids. So far I have used commercial reagents and every time got to see transfection efficiency nearly 70-80%. However, since these...

06 August 2014 3,770 1 View