| Overview | |
|---|---|
| Description | Creative Biolabs offers a wide range of reporter bioassays kits to characterize and develop novel monoclonal antibody (mAb) and cell-based therapeutics. The CTLA-4 Blockade Bioassay Kit is a bioluminescent cell-based assay that can be used to measure the potency and stability of antibodies and other biologics targeting CTLA-4. |
| Features | Convenience and minimizes variability; Easily applicable to high-throughput workflows. |
| Applications | To measure the potency and stability of antibodies and other biologics targeting CTLA-4. |
| Application Notes | It is recommended to conduct a preliminary experiment before the formal experiment. |
| Molecular Mechanism of Action | The Bioassay Kit consists of two genetically engineered cell lines, CTLA-4 Effector Cells and aAPC/Raji Cells. When co-cultured, the CTLA-4/CD80 and CD86 interaction inhibits CD28 pathway-activated luminescence. The addition of anti-CTLA-4 antibody blocks the CTLA-4/CD80 and CD86 interaction, thereby re-establishing CD28 pathway-activated luminescence, which can be detected in a dose-dependent manner by addition of Bio-Glo Reagent and quantitation with a luminometer. |
| Qualified With | Prequalified according to ICH guidelines |
| Assay Measures | Uses Bio-Glo Luciferase Assay System |
| Research Areas | Drug discovery; Function assay |
| Specification | |
|---|---|
| Size | 1EA |
| Components | 1 vial x5 CTLA-4 Effector Cells (0.8ml x5) 1 vial x5 aAPC/Raji Cells (0.8ml x5) 36ml x5 RPMI 1640 Medium 4ml x5 Fetal Bovine Serum 1 vial x5 Bio-Glo Luciferase Assay Substrate (lyophilized) 10ml x5 Bio-Glo Luciferase Assay Buffer |
| Target | CTLA-4 |
| Species Reactivity | Human |
| Target/Description | CTLA-4 (Cytotoxic T-lymphocyte-associated protein 4), also known as CD152, is an immune inhibitory receptor constitutively expressed on regulatory T cells (Tregs) and upregulated in activated T cells. CTLA-4 plays a critical role in regulating immune responses to tumor antigens and autoantigens. When CTLA-4 expression is upregulated on the surface of T cells, the T cells bind B7 with a higher avidity, and thus outcompete the positive co-stimulatory signal from CD28. In addition, engagement of CTLA-4 by either of its ligands, CD80 (B7-1) or CD86 (B7-2) on an adjacent antigen presenting cell (APC) inhibits CD28 co-stimulation of T cell activation, cell proliferation and cytokine production. |
| Alternative Names | CD152 |
| UniProt ID | P16410 |
| Compatibility | Each kit contains sufficient reagents for 600 assays using the inner 60 wells of ten 96-well plates which can do 20 antibody analysis (10 concentration dose*3 repeats). |
| Format | 96- and 384-well plate formats |
| Storage | Upon arrival, immediately transfer the cell vials to below -140°C (freezer or liquid nitrogen vapor phase) for long-term storage. Do not store cell vials submerged in liquid nitrogen. Do not store cell vials at -80°C as this will negatively affect cell viability and performance. Store Bio-Glo Luciferase Assay Buffer, Bio-Glo Luciferase Assay Substrate and Fetal Bovine Serum at -20°C. Avoid multiple freeze-thaw cycles of the serum. We recommend preparing Bio-Glo Reagent immediately before use. Do not store and use the reconstituted reagent. Store RPMI 1640 Medium at 4°C protected from fluorescent light. |
| Handling | Please place the product at proper storage condition once received. |
| Shipping | Components are shipped separately by dry ice and wet ice. |
For Research Use Only.
