Splenocyte test with new peptide 3-12-12

2015-01-13

azim58 - Splenocyte test with new peptide 3-12-12


Splenocyte test with new peptide 3-12-12

I now have a new SIINFEKL peptide (part of the ovalbumin sequence). In
the last ELISPOT I performed with Shen with the old peptide, the
splenocytes with the target cells with no peptide exhibited more of a
response than the splenocytes with target cells with peptide. I
hypothesized that perhaps the peptide was harming my T cells somehow.
Perhaps the DMSO used to dissolve the peptide was harming the cells. I
plan to obtain some splenocytes, perform a trypan blue cell count,
incubate the cells with peptide, and perform another trypan blue cell
count. If it appears that the peptide is not killing my cells, I will
perform another ELISPOT assay.

How should I dilute my new SIINFEKL peptide (H20, PBS, DMSO . . .?)?
Research about dilution of SIINFEKL peptide 3-12-12


Protocols I plan to follow
cardiac puncture
Surgical Removal of Spleens
Splenocyte Preparation Protocol

Results
Peptide Incubation Test 3-13-12.xlsx

Summary
Ova ELISA Summary 3-18-12.pptx