Absorption of e coli antibodies 5-29-12

2014-08-29

azim58 - Absorption of e coli antibodies 5-29-12

Although we had originally decided that it was not necessary to remove
the E coli, GST, and CpG antibodies, I think I will go ahead and give
this a try since I will be repeating the array experiment with 1 nM of
secondary.

Items
Mus SMC27mer 2 ug/uL 6/7/11 peptide from Shen located at -20 C Tumor cDNA
Library 1-20-11 KW
GST Electro protein 1.18 ug/uL from Shen located at -20 C Tumor cDNA
Library 1-20-11 KW
CpG ODN 2216-3 5 ug/mL 5-21-12 from Shen located at -20 C Tumor cDNA
Library 1-20-11 KW
E coli lysate pET32 TEV Item 9-21-11 KW

I plan to apply 12 1:200 dilutions of mouse anti-SMC1fs antibody to e
coli lysate three times, CpG three times, and GST three times. I will
then take four of these wells to measure the remaining activity against
SMC1fs, e coli lysate, CpG, and GST.

I will use my standard ELISA Protocol
Layout of plate for this experiment:
"L:\storage\CIM Research Folder\DR\2012\5-29-12\e coli ab removal
5-29-12.xlsx"

I will coat everything at 1 ug/mL, and use 1:200 dilutions for everything.

===========================================================================
After 3 absorption steps for all 3 antigens, I measured the reactivity
for the 3 antigens and SMC1fs.
Raw Data Location
S:\Research\Cancer_Eradication\Users\kwhittem\kwhittem\Raw Data Often
Originally on Research Drive\2012\5-31-12 ELISA
Graph of the four wells
"L:\storage\CIM Research Folder\DR\2012\5-31-12\ELISA of absorbed sera
5-31-12.xlsx"

Note that I probably would have obtained better absorption of the
antibodies if I coated at 4 C overnight rather than 1 hr at 37 C.

Item: Mouse anti-SMC1fs E coli, GST, CpG abs absorbed located at tumor
cDNA expression library 1-20-11 -20 C

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