Bio-Rad ProteinChip Antibody Capture Kit - Manual (Page 3)

© 2007 Bio-Rad Laboratories, Inc.
The key component of this kit is the ProteinChip PG20 array (ProteinChip PS20 array
precoupled with recombinant protein G (Pierce). Any antibody capable of binding
protein G can be used with the kit (see appendix for details). The kit also contains
positive (TNF-
α antigen-antibody pair) and negative (bovine IgG) controls and a
crosslinking reagent to bind antibodies covalently to ProteinChip PG20 arrays.
Materials
Materials Included
n
TNF-
α antibody (0.2 mg/ml anti-human TNF-α goat polyclonal antibody in
phosphate buffered saline (PBS), 20 µl
n
TNF-
α antigen* (1.0 µM recombinant human TNF-α antigen in 0.1% bovine
serum albumin (BSA)/PBS), 10 µl
n
Bovine IgG (lyophilized), 750 µg
n
Wash buffer (0.5% Triton X-100 in PBS); 2 bottles, 120 ml each
n
PBS; 2 bottles, 120 ml each
n
1x PBS; 1 bottle, 4 ml**
n
ProteinChip PG20 arrays, 6
n
Bis(sulfosuccinimidyl) suberate (BS 3 ) crosslinking reagent (0.5 mg/vial), 2
n
Deactivation buffer (0.5 M ethanolamine in PBS, pH 8.0), 4 ml
n
ProteinChip sinapinic acid (SPA) energy absorbing molecules (EAMs), 5 mg
n
Instruction manual
* Contains equimolar amounts of TNF with and without N-terminal methionine, yielding a double peak.
** The 4 ml PBS should be used to dilute the bovine IgG, antibody, and antigen, reconstitute the
crosslinking reagent, and deposit to the spots of the ProteinChip array when needed.