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