Daniel:Notebook/ComboLock/2016-5-26
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Antibody Prep
I found some columns Dinh had that were the product I've been waiting on, so I am going to use those to test out the antibody linkage and binding protocols. I'll use the VIM protein again, since it has the highest expression.
Protocol
- Antibody Activation
- Resuspend VIM with 100 uL PBS (final conc 1 mg/mL)
- Equilibrate a 40 kDa spin column 4 times with 200 uL PBS, (100 mM pH 7.3)
- Spin at 1000xg for 2 min
- Add 20 uL antibody to the column and spin for 3 min at 1000xg; collect in a new tube
- Add 2 uL sulfo-SMCC to sample
- Incubate for 2 hr at 4C with 3 times intermittent mixing
- Equilibrate a new 40 kDa column with 100 mM phosphate buffer with 20 mM EDTA
- Transfer antibodies to new column and spin for 3 min at 1000xg
- Oligonucleotide Activation