Intended use

This immunoassay kit allows for the use in vitro quantitative determination of Mouse Granulocyte-Macrophage Colony Stimulating Factor, GM-CSF concentrations in cell culture supernates, serum, plasma and other biological fluids.

 

Introduction

Granulocyte-macrophage colony stimulating factor (GM-CSF) was initially identified by its ability to stimulate the formation of granulocyte and macrophage colonies from hematopoietic progenitor cells in semi-solid media. In vivoand in vitro studies have now demonstrated that GM-CSF is a pleiotropic cytokine that induces myeloid precursor cells to proliferate and differentiate into neutrophils, monocyte/macrophages and eosinophils. GM-CSF has also been shown to stimulate the proliferation and differentiation of erythroid and megakaryocyte progenitor cells. In addition, GM-CSF is a survival factor for and activates the effector functions of mature neutrophils, monocytes/macrophages and eosinophils. GM-CSF is produced by a number of different cell types (including activated T cells, B cells, macrophages, mast cells, endothelial cells, fibroblasts, synoviocytes and various tumor cells) in response to cytokines or immune and inflammatory stimuli.

 

Test principle

The microtiter plate provided in this kit has been pre-coated with an antibody specific to GM-CSF. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for GM-CSF and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB  substrate solution is added to each well. Only those wells that contain GM-CSF, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of **0 nm ± 2 nm. The concentration of GM-CSF in the samples is then determined by comparing the O.D. of the samples to the standard curve.