The CD28:B7-1[Biotinylated] Inhibitor Screening Assay Kit is designed for screening and profiling inhibitors of CD28:B7-1 signaling. The key to this kit is the high sensitivity of detection of biotin-labeled B7-1 by streptavidin-HRP. Only a few simple steps on a microtiter plate are required for the assay. First, CD28 is coated on a 96-well plate. Next, B7-1 is incubated with CD28 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.
This kit comes in a convenient 96-well format, with biotin-labeled B7-1, purified CD28, streptavidin-labeled HRP, and assay buffer for 100 binding reactions.
Materials Required But Not Supplied
PBS (Phosphate buffered saline) Luminometer or microplate reader capable of reading chemiluminescence Rotating or rocker platform
HRP chemiluminescent substrate A (transparent bottle)
6 ml
+4°C
HRP chemiluminescent substrate B (brown bottle)
6 ml
+4°C
79699
White 96-well microplate
1
+4°C
UniProt #
P10747
Background
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. CD28:B7-1 interaction is an important drug target for the regulation of T cells involved in autoimmunity, inflammation, tumor recognition, and immune tolerance.
References
1. Keir, M.E., et al. Immunol. Rev. 2005, 204: 128-143. 2. Haas, C., et al. Int. J. Cancer. 2006, 1;118: 658-667.
Storage/Stability
Stable at least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Instructions for Use
See assay kit data sheet for detailed protocol
Applications
This kit is useful for screening for inhibitors of B7-1 binding to CD28.
Shipping Temperature
-80°C (dry ice)
Contraindications
DMSO >1%, strong acids or bases, ionic detergents, high salt.
Warnings
Avoid freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.