Absorption+of+e+coli+antibodies+5-17-12

I would like to absorb the e coli, GST, and CpG antibodies from the SMC1fs sera (see SMC1fs Immunization Timeline) (note that alum was also used as an adjuvant, but I do not think it is possible to raise any antibodies against alum which is just a type of aluminum hydroxide (KAl(SO4)2·12H2O)), and I would like to absorb the e coli antibodies from the 4T1 and naive sera. Then I would like to reapply this sera to the nitrocellulose tumor cDNA expression library array.

Before absorbing any antibodies, I will do an ELISA to measure the antibody response to start with. Then I will absorb the antibodies. Then I will check the effectiveness of the absorption by doing another ELISA. see ELISA Protocol

After the primary is incubated for 1 hr, I will transfer the primary to a new plate coated completely with SMC1fs to see how much SMC1fs antibody was removed by the various incubations. I need to figure out which e coli lysate I will use.

E coli lysate pET32 TEV Item 9-21-11 KW E coli SlyD Extract Item

5-18-12 Coated ELISA plate at 4 C over weekend. Layout of ELISA plate: "L:\storage\CIM Research Folder\DR\2012\5-17-12\test of smc1fs sera 5-17-12.xlsx" I also coated another plate that was completely coated with GST-SMC1fs protein.

Items in this experiment E coli lysate item at 4 C in 15 mL tube CpG item at 4 C at 4 C in 15 mL tube GST electro item at 4 C in 15 mL tube Mus SMC27mer item at 4 C in 15 mL tube

5-21-12 I used the wrong kind of plate (round bottom assay plate), and I will need to repeat the experiment. I should have used a flat bottom Maxisorb Nunc plate.

New layout of ELISA plate "L:\storage\CIM Research Folder\DR\2012\5-21-12\test of smc1fs sera 5-21-12.xlsx"

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

Data from experiment "S:\Research\Cancer_Eradication\Users\kwhittem\kwhittem\Raw Data Often Originally on Research Drive\2012\5-22-12 ELISA\5-22-12 ELISA.xlsx"

Summary of experiment "S:\Research\Cancer_Eradication\Users\kwhittem\kwhittem\Raw Data Often Originally on Research Drive\2012\5-22-12 ELISA\Summary of Mouse Sera ELISA 5-22-12.docx"

Next experiment related to this one Absorption of e coli antibodies 5-25-12