CD155:CD96 [Biotinylated] Inhibitor Screening Chemiluminescence Assay Kit
The CD155:CD96 [Biotinylated] Inhibitor Screening Chemiluminescence Assay Kit is an ELISA-based assay designed for screening and profiling molecules that block the binding between CD155 (cluster of differentiation 155) and CD96 (cluster of differentiation 96). This kit comes in a convenient 96-well format, with enough recombinant human biotin-labeled CD96 (amino acids 22-503), CD155 (amino acids 27-343), streptavidin-labeled HRP, and assay buffer for 100 reactions.
Figure 1: Illustration of the mechanism of CD155: CD96 [Biotinylated] Inhibitor Screening Chemiluminescence Assay Kit.
A 96-well plate is coated with CD155 protein. After blocking, the plate is pre-incubated with an inhibitor or neutralizing antibody. After incubation with Biotin-CD96, the plate is washed and Streptavidin-HRP is added. The ELISA ECL substrate is added, and the resulting signal can be measured using a chemiluminescence microplate reader. The chemiluminescence signal is proportional to the binding of CD155 to CD96.
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- 1x PBS Buffer (Phosphate Buffer Saline, pH 7.4)
- PBS-T Buffer (1x PBS buffer with 0.05% Tween-20)
- Microplate reader capable of reading chemiluminescence
- Adjustable micropipettor and sterile tips
- Rotating or rocker platform
| Catalog # | Name | Amount | Storage |
| CD96, His-Tag, Avi-Tag, Biotin-Labeled* | 10 µg | -80°C | |
| 79063 | CD155, Fc-Avi-Tag* | 2 x 25 µg | -80°C |
| 5x PP-02 Buffer | 4 ml | -20°C | |
| 79728 | Blocking Buffer 2 | 40 ml | +4°C |
| 79742 | Streptavidin-HRP | 10 µl | +4°C |
| 79670 | ELISA ECL Substrates A (translucent bottle) | 6 ml | Room Temp |
| ELISA ECL Substrates B (brown bottle) | 6 ml | Room Temp | |
| 79699 | White 96-well microplate | 1 | Room Temp |
* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.
CD96, also known as Tactile (T cell activation, increased late expression), is an immune checkpoint receptor of the immunoglobulin superfamily of proteins found in T and NK cells. Other proteins of the same superfamily include CD226 (also known as DNAM-1) and TIGIT (T cell immunoglobulin and ITM domain), with CD96 and TIGIT acting as inhibitory receptors and CD226 as a co-stimulatory receptor. CD96 binds to CD155, competing with CD226. CD96 plays a role in NK-cell adhesion to target cells, inhibiting NK cell cytokine release. It also plays an inhibitory role in Th9 cells and is a cancer stem cell marker. An imbalance in CD155/CD96 and CD155/CD226 can lead to lack of or to excessive immune responses. In LUAD (lung adenocarcinoma) the levels of CD96 in CD8+ T cells are elevated compared to healthy individuals, possibly promoting a hypo-immune response. CD96, together with TIGIT, has become a widely researched target in immunotherapy, and blockage of CD96 binding to CD155 combined with other therapies, such as anti-PD-1 (programmed death protein 1) or doxorubicin chemotherapy, has shown great promise. The development of strategies to block CD96 inhibitory immune responses will continue to be an active area de research, opening new avenues of treatment for cancer patients.
Zhang H., et al., 2020 Cellular & Molecular Immunology 18:1575-1577.
Liu F., et al., 2020 Scientific Reports 10:10768.
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