Active Macrophage Inflammatory Protein 1 Beta (MIP1b)

Macrophage Inflammatory Protein 1 Beta (MIP‑1β, A093Hu), also known as CCL4, is a small pro‑inflammatory CC chemokine mainly secreted by macrophages, monocytes, dendritic cells and activated lymphocytes. It participates in immune surveillance, inflammatory response and host defense against pathogens by recruiting and activating immune cells. MIP‑1β is widely involved in infectious diseases, autoimmune disorders and tumor‑related immune infiltration. It binds to the chemokine receptor CCR7 (B647Mu) with high affinity, mediating immune cell chemotaxis and migration. Briefly, CCR7 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μL were then transferred to MIP‑1β-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CCR7 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630 nm immediately. Measured by its binding ability in a functional ELISA. When recombinant human MIP‑1β is lmmobilized at 2 µg/mL(100 µL well), the concentration of CCR7 that produces 50% optimal bindingresponse is found to be approximately 0.154 µg/mL.