Description
Principle of the Assay: This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-M-CSF polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-M-CSF polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the M-CSF amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of M-CSF can be calculated.
Background: Macrophage colony-stimulating factor, or M-CSF, is a secreted cytokine which influences hematopoietic stem cells to differentiate into macrophages or other related cell types. Ladner et al. (1987) showed that there are 2 forms of M-CSF, with 224 and 522 amino acids, resulting from alternative splicing. It is a hematopoietic growth factor that is involved in the proliferation, differentiation, and survival of monocytes, macrophages, and bone marrow progenitor cells. It released by osteoblasts (as a result of endocrine stimulation by parathyroid hormone) exerts paracrine effects on osteoclasts. M-CSF binds to receptors on osteoclasts inducing differentiation, and ultimately leading to increased plasma calcium levels—through the resorption (breakdown) of bone. More recently, it was discovered that CSF-1 and its receptor CSF1R are implicated in the mammary gland during normal development and neoplastic growth