CTLA4 [Biotinylated]:B7-1 Inhibitor Screening Assay Kit
The activation of naïve T cells requires two signals, the specific T cell receptor recognition of MHC/Antigen on the surface of the antigen-presenting cell (APC), and the binding of B7-1 (CD80) ligand on the APC with the CD28 receptor on the T cell surface. Conversely, binding of CTLA4 to B7-1 on the T-cell surface results in an inhibitory signal and prevents T-cell activation. CTLA4B7-1 interaction is an important drug target for the regulation of the host’s response to cancer. The CTLA4 [Biotinylated]:B7-1 Inhibitor Screening Assay Kit is designed for screening and profiling inhibitors of CTLA4:B7-1 signaling. The key to this kit is the high sensitivity of detection of biotin-labeled CTLA4 by streptavidin-HRP. Only a few simple steps on a microtiter plate are required for the assay. First, B7-1 is coated on a 96-well plate. Next, CTLA4[B] is incubated with B7-1 on the plate. Finally, the plate is treated with streptavidin-HRP followed by addition of an HRP substrate to produce chemiluminescence, which can be measured using a chemiluminescence reader.
PBS (Phosphate buffered saline)
Luminometer or fluorescent microplate reader capable of reading chemiluminescence
Rotating or rocker platform
| Catalog # | Component | Amount | Storage | |
| 71125 | B7-1 | 10 µg | -80°C | Avoid freeze/ thaw cycles! |
| 71152 | CTLA4, Biotin-labeled | >1 µg | -80°C | |
| 79742 | Streptavidin-HRP | 10 µl | +4°C | |
| 79311 | 3x Immuno Buffer 1 | 50 ml | -20°C | |
| 79728 | Blocking Buffer 2 | 50 ml | +4°C | |
| 79670 | ELISA ECL substrate A (transparent bottle) |
6 ml | Room Temp |
|
| ELISA ECL substrate B (brown bottle) |
6 ml | Room Temp |
||
| 79699 | White 96-well microplate | 1 | +4°C | |
1. Linsley, P.S., et al. J. Exp. Med. 1991, 174(3): 561-569.
2. Hurwitz, A.A., et al. Canc. Res. 2000, 60: 2444-2448.
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