Description
system_update_altDatasheet
| Product Name: | GeniePlex Human Immune Checkpoint Multiplex Assay (10-Plex) (96 Tests) |
| Product Code: | HUAMPM041 |
| Size: | 96 Tests |
| Reactivity: | Human |
| Analytes (10): | sCTLA-4/sCD152, sTIM-3/sHAVCR2, OX40/TNFRSF4/sCD134, sB7-H3/sCD276, 4-1BB/TNFRSF9/sCD137, PD-L1/sCD274/sB7-H1, sLAG-3/sCD223, CD279/sPD-1, GITR/TNFRSF18/sCD357, and sICOS/sCD278 |
| Sensitivity (LODs): | sCTLA-4/sCD152: N/A pg/mL, sTIM-3/sHAVCR2: N/A pg/mL, OX40/TNFRSF4/sCD134: N/A pg/mL, sB7-H3/sCD276: N/A pg/mL, 4-1BB/TNFRSF9/sCD137: < 5 pg/mL, PD-L1/sCD274/sB7-H1: < 20 pg/mL, sLAG-3/sCD223: < 5 pg/mL, CD279/sPD-1: < 20 pg/mL, GITR/TNFRSF18/sCD357: < 20 pg/mL, sICOS/sCD278: < 10 pg/mL |
| Sample types: | Cell culture supernatant, serum, plasma, bodily fluid and tissue/cell lysate |
| Assay Type: | Multiplex |
| Detection Method: | Flow Cytometry |
| Assay Time: | 2 hours |
| 1x Ab-conjugated beads (premixed): | anti-human sCTLA-4/sCD152, anti-human sTIM-3/sHAVCR2, anti-human OX40/TNFRSF4/sCD134, anti-human sB7-H3/sCD276, S4P4 - anti-human 4-1BB/TNFRSF9/sCD137, S4P8 - anti-human PD-L1/sCD274/sB7-H1, S4P9 - anti-human sLAG-3/sCD223, S5P8 - anti-human CD279/sPD-1, S5P8 - anti-human GITR/TNFRSF18/sCD357, S5P9 - anti-human sICOS/sCD278 |
| 2x Biotin-detection Ab (premixed): | Biotin - anti-human sCTLA-4/sCD152, Biotin - anti-human sTIM-3/sHAVCR2, Biotin - anti-human OX40/TNFRSF4/sCD134, Biotin - anti-human sB7-H3/sCD276, Biotin - anti-human 4-1BB/TNFRSF9/sCD137, Biotin - anti-human PD-L1/sCD274/sB7-H1, Biotin - anti-human sLAG-3/sCD223, Biotin - anti-human CD279/sPD-1, Biotin - anti-human GITR/TNFRSF18/sCD357, Biotin - anti-human sICOS/sCD278 |
| Buffers: | 2x NR dAb Diluent. One vial containing 1.5 mL of biotin detection antibody diluent. |
| Lyophilized Standard Mix: | Lyophilized Standard Mix- Human Immune Checkpoint 10-Plex. Vials A, B and C containing lyophilized recombinant human sCTLA-4/sCD152, sTIM-3/sHAVCR2, OX40/TNFRSF4/sCD134, sB7-H3/sCD276, 4-1BB/TNFRSF9/sCD137, PD-L1/sCD274/sB7-H1, sLAG-3/sCD223, CD279/sPD-1, GITR/TNFRSF18/sCD357, sICOS/sCD278. |
| LLOQs: | sCTLA-4/sCD152: N/A pg/mL, sTIM-3/sHAVCR2: N/A pg/mL, OX40/TNFRSF4/sCD134: N/A pg/mL, sB7-H3/sCD276: N/A pg/mL, 4-1BB/TNFRSF9/sCD137: < 10 pg/mL, PD-L1/sCD274/sB7-H1: < 50 pg/mL, sLAG-3/sCD223: < 10 pg/mL, CD279/sPD-1: < 50 pg/mL, GITR/TNFRSF18/sCD357: < 50 pg/mL, sICOS/sCD278: < 20 pg/mL. |
| ULOQs: | sCTLA-4/sCD152: N/A pg/mL, sTIM-3/sHAVCR2: N/A pg/mL, OX40/TNFRSF4/sCD134: N/A pg/mL, sB7-H3/sCD276: N/A pg/mL, 4-1BB/TNFRSF9/sCD137: > 5,000 pg/mL, PD-L1/sCD274/sB7-H1: > 10,000 pg/mL, sLAG-3/sCD223: > 5,000 pg/mL, CD279/sPD-1: > 10,000 pg/mL, GITR/TNFRSF18/sCD357: > 20,000 pg/mL, sICOS/sCD278: > 10,000 pg/mL. |
| Standard dose recovery: | 70-130% |
| Intra-assay CV: | Intra-assay CV: <10% |
| Inter-assay CV: | Inter-assay CV: <20% |
| Cross-reactivity of analytes in the Panel: | Negligible |
| Sample volume: | 15 µL/test |
*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
A filter plate washer is required for the following protocol. For more information on GeniePlex click here.
| Step | Protocol |
| 1. | Prepare the filter plate template. |
| 2. | Vortex working bead suspension for 15 seconds. |
| 3. | Add 45 µL of capture bead working suspension to each well. |
| 4. | Remove buffer in the wells by using the Filter Plate Washer connected to a vacuum source. |
| 5. | Remove buffer in the wells by using the Filter Plate Washer connected to a vacuum source. |
| 6. | Add 30 µL of CCS, SPB or TL Assay Buffer to each sample well. |
| 7. | Add 15 µL of samples to each sample well. Add 45 µL of standards to each standard well. Cover the plate with a plate seal. |
| 8. | Incubate on the shaker (set at 700 rpm) for 60 min at room temperature. Protect from light. |
| 9. | Remove the plate seal. Remove solutions in the wells by using the Filter Plate Washer. |
| 10. | Remove solutions in the wells by using the Filter Plate Washer. |
| 11. | Wash the wells three times with 100µL 1x Wash Buffer. |
| 12. | Gently tap the plate bottom onto paper towels to remove residual buffer. |
| 13. | Add 25µL of biotinylated antibody working solution to each well. Cover with plate seal. |
| 14. | Incubate on the shaker (700 rpm) for 30 min at room temperature. Protect from light. |
| 15. | Remove plate seal. Remove solutions. Wash three times with 100 µL 1x Wash Buffer. |
| 16. | Gently tap the plate bottom onto paper towels. |
| 17. | Add 25µL of streptavidin-PE working solution to each well. Cover with plate seal. |
| 18. | Incubate on the shaker (700 rpm) for 20 min at room temperature. Protect from light. |
| 19. | Remove plate seal. Remove solutions. Wash twice with 100 µL 1x Wash Buffer. |
| 20. | Add 150-300µL of 1x Reading Buffer to each well to re-suspend beads. Cover with plate seal. |
| 21. | Shake for 30 seconds at 700 rpm. Read on a flow cytometer. |
GeniePlex Technology
GeniePlex uses a unique mix of antibody-coated encoded microparticles providing an ultra-sensitive technology for the quantitation of analytes.
