CD112R:CD112[Biotinylated] Inhibitor Screening Assay Kit
The CD112R:CD112 [Biotinylated] Inhibitor Screening Colorimetric Assay Kit is designed for screening and profiling of inhibitors or neutralizing antibodies able to block the interaction between CD112R (cluster of differentiation 112 receptor) and CD112. This kit comes in a convenient 96-well format, with enough biotin-labeled CD112 (amino acids 32-360), purified CD112R (amino acids 100-330), streptavidin-labeled HRP, and assay buffer for 100 binding reactions.
Figure 1: CD112R:CD112 [Biotinylated] Inhibitor Screening Colorimetric Assay Kit schematic.
The key to this kit is the high sensitivity of detection of biotin-labeled CD112 by streptavidin-HRP. CD112R is coated on a 96-well plate. After blocking, the protein is pre-incubated with the inhibitor or neutralizing antibody. Upon subsequent incubation with Biotin-CD112, the plate is treated with Streptavidin-HRP followed by addition of a colorimetric HRP substrate to produce color, which can be quenched and measured using a UV/Vis microplate reader. Absorbance values are proportional to the binding of CD112R to CD112.
- PBS (Phosphate Buffered Saline)
- 1N HCl (aqueous)
- Rotating or rocker platform
- UV/Vis spectrophotometer microplate reader capable of reading absorbance at λ=450 nm
| Catalog # | Name | Amount | Storage |
| 79116 | CD112R (PVRIG), Fc-Fusion, Avi-Tag (Human) HiP™* | 10 µg | -80°C |
| 72231 | CD112 (PVRL2), His Avi-Tag, Biotin-labeled (Human) HiP™* | 5 µg | -80°C |
| 79311 | 3X Immuno Buffer 1 | 50 ml | -20°C |
| 79728 | Blocking Buffer 2 | 50 ml | +4°C |
| 79742 | Streptavidin-HRP | 10 µl | +4°C |
| 79651 | HRP Colorimetric Substrate | 6 ml | +4°C |
| 79964 | Clear 96-well microplate | 1 | Room Temp |
*The initial concentration of both proteins is lot-specific and will be indicated on the tube containing the protein.
CD112 (Poliovirus receptor-related 2, PVRL2), widely expressed on antigen presenting cells and tumor cells, is the high affinity ligand of CD112R (Poliovirus receptor related immunoglobulin domain containing, PVRIG). CD112R is found in NK and T cells, and in T cells it is found particularly in CD8+ T cells. High levels of this protein are found in NK, CD8+ and CD4+ T cells of patients with kidney, ovary, lung, prostrate and endometrium cancer and acute myeloid leukemia (AML). CD112 has also been linked to tumor angiogenesis, growth and metastasis. It has a wider pattern of expression, being also present in endothelial cells, neurons and fibroblasts. CD112-CD112R interaction is a positive immune checkpoint that enhances human T cell response and has emerged as an attractive therapeutic target for oncology. TIGIT (T cell immunoreceptor with Ig and ITIM domains) and CD226 (also called DNAM-1, DNAX Accessory Molecule-1) also bind CD112. The development of inhibitors able to prevent the interaction between CD112R-CD112 alone or in combination with blockage of TIGIT-CD155/CD112 interaction may prove beneficial in cancer therapy.
Zhu, Y., et al. J. Exp. Med. 2016, 213(2):167-76
Torphy, R., et al. Int. J. Mol. Sci. 2017, 18(12): E2642
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