Human MyD88 ELISA Kit

A high-quality sandwich ELISA from Assay Genie designed for the quantitative measurement of human MyD88 (also known by the synonyms MYD88 and MYD88D, myeloid differentiation primary response protein MyD88) in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Reagents should be stored at 2-8°C, where they remain stable for up to 12 months.

The assay protocol involves six main steps: equilibrate reagents and the TMB substrate to room temperature; incubate standards, samples and controls on the pre-coated plate at 37°C for 90 minutes to allow antigen binding; add a biotin-labelled detection antibody and incubate at 37°C for 60 minutes; add HRP-Streptavidin (SABC) and incubate at 37°C for 30 minutes; develop colour by adding TMB substrate and incubating in the dark for 10-20 minutes; and stop the reaction, then read absorbance at 450 nm immediately.

Sample preparation guidance includes allowing blood to clot and centrifuging serum at 1000 × g for 20 minutes; collecting plasma in anticoagulant tubes and centrifuging at 1000 × g for 15 minutes at 2-8°C; homogenising tissue in PBS with protease inhibitors; centrifuging cell culture supernatant at 2500 rpm for 5 minutes; and lysing cells with lysis buffer containing protease inhibitors. Linearity has been validated across serum (91-104% at 1:2, 83-99% at 1:4, 83-96% at 1:8), EDTA plasma (90-103% at 1:2, 91-101% at 1:4, 80-95% at 1:8) and heparin plasma (85-105% at 1:2, 84-101% at 1:4, 80-92% at 1:8). Recovery ranges are 86-103% (average 94%) for serum, 85-103% (average 94%) for EDTA plasma, and 85-104% (average 93%) for heparin plasma. MyD88 is a key adaptor protein in innate immune signalling, and this kit supports research applications in inflammation, autoimmune disorders, infectious disease and cancer.