Mouse CD274/B7-H1/PD-L1 ELISA Kit

A Sandwich ELISA kit from Assay Genie for the quantitative measurement of mouse CD274 (also known as B7-H1 or PD-L1), a key immune checkpoint protein involved in regulating immune responses and maintaining immune homeostasis. The kit is designed for serum, plasma, cell culture supernatants, cell or tissue lysates, and other liquid samples, and is intended for immunology research into conditions such as autoimmune disease, infectious disease, and cancer, where dysregulated CD274 expression has been reported.

According to the manufacturer, the assay uses a double-antibody sandwich ELISA format with colorimetric detection. Sensitivity is 9.375 pg/mL and the standard range is 15.625–1000 pg/mL. Reported linearity across 1:2, 1:4, and 1:8 dilutions (n=5) is 88–98%, 87–104%, and 85–99% for serum; 87–101%, 92–101%, and 94–101% for EDTA plasma; and 80–96%, 82–100%, and 94–100% for heparin plasma. Recovery ranges (with average) are 85–104% (95%) for serum, 94–105% (100%) for EDTA plasma, and 85–104% (98%) for heparin plasma. The kit is stored at 2–8°C for 12 months.

The protocol comprises six steps: reagent and TMB substrate equilibration to room temperature; primary incubation at 37°C for 90 minutes; biotin-labeled detection antibody binding at 37°C for 60 minutes; HRP-Streptavidin incubation at 37°C for 30 minutes; TMB substrate color development in the dark for 10–20 minutes; and stop reaction followed by absorbance reading at 450 nm. Sample preparation guidance includes blood clotting and centrifugation at 1000 × g for 20 minutes for serum, anticoagulant collection and centrifugation at 1000 × g for 15 minutes at 2–8°C for plasma, homogenization in PBS with protease inhibitors for tissue, and lysis buffer with protease inhibitors for cell lysates.