Sudip Das

12 Questions 58 Answers 0 Followers

Questions related from Sudip Das

Chemical fixation vs heat killing of bacteria: which method is better to preserve MAMPs?

Hello Scientific World, I am working on host-microbe interaction specifically bacterial stimulation of macrophages. In my in vitro experiments, I would like to use dead but intact bacteria to...

06 June 2018 7,646 2 View

What is an accepted equivalent of non-parametric version two-way ANOVA in R package?

I use R for my statistical analysis. I want to analyse a kinetics experiment that involves measurement over 3 time points with control, genetic background 1, 2 and 3. Can anyone suggest a test and...

10 October 2016 8,585 4 View

How to differentiate HL-60 to Neutrophils in suspension and which markers to check ?

I have started working on HL-60 cell lines. My aim is to look for Calcium flux upon activation, chemotaxis and also infection with bacteria. I followed Dr. Steven Ackerman's protocol for thawing...

05 May 2016 10,051 11 View

Can anyone suggest cell lines to study cytokines production upon infection in vitro ?

I want to study cytokine production upon S. aureus infection in vitro. I have the following concerns: 1. Which human/mouse cells are best suited ? 2. Which assay is best suited in terms of both...

09 September 2015 10,096 5 View

What's the best way to detach primary macrophages from surface of tissue culture plates/dishes ?

I have started working with primary human and murine macrophages and have successfully differentiated and used for infection experiments. I perform mostly CFU assays with bacteria. Although, I...

04 April 2015 3,881 16 View

What are major inflammatory cytokines that can be quantified upon lung infection ?

I am testing a bacterial mutant in pneumonia model and also wanted to check for inflammation. Apart from microscopic examination using HE staining, I wanted to quantify inflammatory cytokines in...

03 March 2014 5,592 6 View

How to get rid of DNA from agar plates?

I want to use mouse tissue homogenates containing bacteria and plate it on agar plates. For my experimental purpose I need to pool the bacteria from the plates. If I scrape off the bacteria, will...

06 June 2013 2,612 2 View

DNA eluted with 6M Guanidine hydrochloride from Streptavidin Magnetic beads, does it need further purification for downstream applications?

1. Will be compatible with Agarose TAE gels, when loaded directly? 2. Will it be compatible in Phusion PCR reactions, further as template? Note: The manufacturer recommends 6m GuHCL for elution...

05 May 2013 8,309 7 View

Which is the best method for elution of DNA (biotin tagged) from Streptavidin Magnetic Beads?

I have primers with Biotin tag. I use them in PCR for enrichment of particular amplicons. I have a feeling that the 6M Guanidine HCL recommended is not doing the best job. I also have to use the...

05 May 2013 5,807 7 View

How to check differentiation efficiency of Human monocyte/macrophage from HL-60 cells?

Hello fellow researchers, Quite recently, we differentiated HL-60 cells into PMNs using DMSO and had a fair amount of success. I usually check for the increase in surface CD11b and also control...

01 January 1970 2,548 5 View

Why does THP-1 cells shown different populations in flow cytometry scatter plot?

Hello Scientific World, I would highly appreciate some help here. I am using THP-1 monocytes as my pet cell line to generate human macrophages. Before every differentiation process, I test my...

01 January 1970 1,465 6 View

How can the expression of Interleukin-10 and MMP12 be stimulated in macrophages ?

Hello, I am currently trying to standardise qPCR primers for IL-10 expression in human macrophages i.e. checking amplification, primer efficiencies. Before the actual stimulation by bacteria, I...

01 January 1970 5,291 4 View