Active Colony Stimulating Factor 1, Macrophage (MCSF)

Colony Stimulating Factor 1 (MCSF), also known as Macrophage Colony-Stimulating Factor (M-CSF), is a crucial hematopoietic growth factor that regulates the survival, proliferation, and differentiation of monocytes and macrophages. It binds to its receptor, MCSFR, playing a fundamental role in innate immunity, bone biology, and homeostasis. Furthermore, MCSF and Interleukin-34 (IL34) are functionally related as they are both high-affinity ligands for the MCSF receptor (MCSFR), activating similar downstream signaling pathways despite their distinct protein structures.Besides,IL34 has been identified as an interactor of MCSF, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human MCSF and recombinant human IL34. Briefly, MCSF was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to IL34-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MCSF 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 450/630nm immediately. The binding activity of recombinant human MCSF and recombinant human IL34 was shown in Figure 1, the EC50 for this effect is 0.0472µg/mL.