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Questions related from Theodore Kapellos
I am doing reverse transcription using a semi-suppressive PCR therefore I introduce a template switching oligo at the 5' end of my transcript which is complementary to the PCR handle at the 3'...
04 October 2018 4,363 2 View
I want to stimulate a murine endothelial cell line (sEnd.1) and test my drugs of interest to quantify anti-inflammatory potential. It is accepted for endothelial cells to be treated with TNF-α...
27 December 2013 1,768 9 View
A question of cytokine kinetics. In an activation assay, we usually want to measure effects of our treatment (whatever this might be) on pro- or anti-inflammatory cytokines. Most commonly...
29 November 2013 2,199 3 View
I homogenized murine lungs in 0.5% w/v HDAB in 50mM sodium phosphate buffer to perform a MPO activity assay. The protocol I do have instructs that I should use 20ul of my samples with 160ul TMB...
02 October 2013 8,478 3 View
I finished an ELISA assay and read it to discover two hours later that I saved the 630nm data which I used to correct my 450nm data. I immediately measured the plate again but I'm not sure whether...
06 September 2013 4,183 21 View
I have some qPCR data which I want to represent in graphs. I used to use the DCt values but the problem is that they are reciprocal to the expression changes (high DCt values means low expression)...
14 July 2013 4,946 2 View
I am stimulating microglial cell lines (BV-2 cells) with LPS and IFN-γ. I want to study how inflammation develops in these cells and therefore want to investigate the pathways activated early in...
05 June 2013 6,410 34 View
I want to inject i.p. Annexin or Dexamethasone to mice for the needs of my in vivo experiments. After these injections I will administer inflammatory mediators and I want to see whether...
20 May 2013 2,875 20 View
I have some primary macrophages and want to make up a phagocytosis assay. It would be very helpful if anyone could share a protocol to get me started.
16 October 2012 8,599 3 View
Short question: why does one add PBS+EDTA in adherent cell cultures to detach them? And is it more preferable than trypsinization?
08 October 2012 7,885 8 View
I have been doing sandwich ELISAs the last week but I ran out of the Costar ELISA plates I had been using and used anoher company's (Greiner One). Both plates were flat bottomed 96 well plates....
23 February 2012 8,881 21 View
