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Questions related from Durgesh Pitale
I want to isolate Leishmania donovani LPG interacting host proteins. Kindly share your references for selecting antibody for the same.
04 January 2018 9,441 1 View
I want to perform Immuno-TEM staining in human blood-derived neutrophils for my antigen. My protein of interest will suppose to be localized on autophagosomes. Please share protocol for the...
18 September 2017 9,087 3 View
I working with human neutrophils. I want to develop phospho mTOR and mTOR western blot for it. Please suggest me protocol for transfer as well as antibody which is suitable for my sample.
06 July 2016 1,826 5 View
I want to study effect of particular protein of PMN on parasite load in mice. In this set of experiment I am planning to adoptively transfer PMNs from wild type mice into parasite infected...
27 February 2016 2,321 1 View
I want to find level of arginase produced in human macrophage. I am referring following protocol. 1) Lysis of human macrophages in lysis buffer (0.1% triton-x-100+ protease inhibitor cocktail) 15...
23 December 2015 4,885 6 View
I am using Phospho-SAPK&JNK pThr183+Tyr185 Antibody (E.665.10) of Thermofisher (Pierce antibody). Protocol followed by me is as follows. 1. PMN fixation with 4% PFA/ 30 min/ 4 degree...
02 December 2015 9,734 3 View
I am expressing Leishmania protein in bacterial system. This sequence has four rare codons of proline, isoleucine, leucine and arginine. I have tried BL-21 DE3, BL-21 Gold, BL-21 RIL, BL-21 RP,...
15 February 2015 5,952 5 View
I am doing ICC staining for endonuclease G in leishmania species. I have not encountered any antibody raised against peptide which has homology with leishmania endonuclease G protein sequence.
05 December 2014 9,258 3 View
I am staining for protein x in leishmania. I have tried so many different method of fixation e.g. 1-4%PFA (10 min), methanol, formaldehyde (1-4% for 5-10min), Acetone:methanol (1:1) in all...
20 November 2014 5,839 3 View
I want to detect ROS and NO generation in primary macrophages. DCF-DA and DAF-FM are the methods used by my lab for ROS and NO detection by Fluorimetric analysis. They do this by trypsinizing the...
01 September 2014 7,933 7 View
I have tried mitotracker staining of leishmania. It is giving very nice staining when parasites are live. When I fixed leishmania with 4% Paraformaldehyde, it looses its mitotracker staining. Also...
27 August 2014 8,688 7 View
I have tried to stain Leishmania mitochondria with a mitotracker but I am not getting clean staining for mitochondria. I have uses the following protocol for staining: 1) 1 micro liter of 100...
18 June 2014 9,240 2 View
I have infected macrophage cell line (THP-1) with Leishmania parasite and now I want entrapped parasite to be used for culturing. How do I let the parasite free from macrophages without harming...
17 May 2014 7,811 2 View
I want to stain for neutrophil granules. Is their any other stain or dye used to stain specifically neutrophils granules?
28 February 2014 526 3 View
I have isolated human neutrophils (from blood) in the inactivated condition for parasite infection study. I need to fix the cells for staining protocol. I have used 1% and 4% paraformaldehyde. I...
06 February 2014 5,920 2 View
I have isolated neutrophils in inactivated state as per experimental need. I am searching for neutrophil cryopreservation protocol. I have used FBS+5%DMSO for cryopreservation. But after revival...
23 January 2014 1,003 2 View
I have obtained nature protocol for isolation on pure neutrophil population. I want to do work on infections response in neutrophil but shucked at neutrophil culture.
29 December 2013 7,635 25 View
