Alok Kumar Singh

9 Questions 18 Answers 0 Followers

Questions related from Alok Kumar Singh

Immunoprecipitated protein band shifts downwards in Western blot using Dynabead protein G ?

I have been using Dynabead protein-G for immunoprecipitation of my target protein in a mix of antigen from two different protein samples. My target protein X is supposed to bind to the protein Y...

21 November 2016 6,763 4 View

Looking for R5 HIV pro viral plasmids?

These plasmids are accessible through AIDS Reagent Program. These are: 1. pNL4-3 and 2. HIV-1 HXB3/BaL Infectious Molecular Clone (pWT/BaL). I assume that these plasmid encoded viruses will be...

22 September 2016 4,359 3 View

Can we use essential and non-essential amino acids in RPMI media used for differentiation of human monocyte-derived macrophages?

Any suggestion that the amino acids will lead to any detrimental change on differentiation of monocytes in to macrophages in conjunction with recombinant M-CSF?

11 July 2016 4,533 3 View

What the the best method of determine the replication of R5 HIV-1 viruses in human macrophages?

I am trying to optimize Taqman based qRT-PCR and p24 based ELISA to determine the viral titer for HIV-1 in macrophages exposed to HIV-1 BaL. Is there any established protocol for TCID determination?

11 July 2016 10,062 2 View

What is the best viral dose, method protocol and time period to investigate the HIV-1BaL (R5 virus) multiplication in PMA activated THP-1 cells?

I have been trying to optimize the viral transduction protocol macrophage like THP-1 cells (PMA activated). I am not sure of the time period for which I should allow the virus exposure to cells?...

11 July 2016 1,277 3 View

What is the best method of purification of HIV-1 viruses?

What is the best method of enrichment and purification of HIV-1 from culture supernatant? Ultracentrifugation or using cut off columns? Is it better to store these viruses in 20% sucrose or 30%...

11 July 2016 2,955 4 View

Can anyone explain why the Beclin-1 level is similar in control and autophagy inducer treated samples?

In a mammalian cell line, autophagy is induced with respect to control cells, still the level of beclin-1 is very similar in both conditions, where a particular inducer of autophagy was used to...

15 July 2014 7,838 2 View

Can we have a high-throughput method of prediction of protein targets for Leishmania GP63?

Leishmania GP63 has been on of the prime proteolytic virulence factor that is known to targets a wide range of substrates that include several phosphatases that are crucial in MAPK signalling and...

07 June 2014 324 0 View

Can we use RIPA buffer as protein lysis in quantitative proteomics experiment wherein iTRAQ labeling is to be done?

I have been regularly using SILAC membrane lysis buffer regularly to perform cell lysis in the iTRAQ based quantitative proteomic experiment. I wish to use RIPA buffer and I don't find any...

12 May 2014 7,171 4 View