Absorption of e coli antibodies 5-17-12

2014-08-29

azim58 - 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.

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

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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

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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"

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

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