Detection Principle:

The double antibody sandwich ELISA method was used in this experiment. Anti human G-CSF monoclonal antibody is coated on the microplate. The G-CSF in the sample and standard will bind with the antibody fixed on the plate, and the free components will be washed away; Horseradish peroxidase labeled anti human G-CSF polyclonal antibody was added, and the unbound antibody was washed away; Add substrate solution (chromogenic agent), and the color of the solution is directly proportional to the bound target protein; Add termination solution; The absorbance was measured with a microplate reader.

Detection Type: Double antibody sandwich method

Form: pre coated 96 well plate

Detection Sample Type: cell supernatant, serum, Plasma

Loading Amount: 100ul

Kit Components:

Precoated 96 well plate, standard, anti human G-CSF detection antibody, dilution buffer, chromogenic solution (a, b), washing solution, termination solution, sa-hrp, plate sealing membrane and instructions.

Sensitivity: 0.089-0.740 pg/ml

Detection Range: 31.3 - 2000 pg/ml

Recovery Range: 96-103%

Storage Method: 2-8 ℃

Standard Curve

Background:

Granulocyte colony stimulating factor (G-CSF) is a 24-25 kDa monomeric glycoprotein that regulates the proliferation, differentiation, and activation of hematopoietic cells in the neutrophil lineage. Mature human G-CSF is a 178 amino acid (AA)Of O-glycosylated proteins, containing two intrachain disulfide bonds. In humans, alternative splicing generates a second isoform with a 3-amino acid deletion. Mouse and human G-CSF share 76% amino acid sequence identity, and the two proteins show species cross reactivity. G-CSF is produced by activated monocytes and macrophages, fibroblasts, endothelial cells, astrocytes, neurons, and bone marrow stromal cells. In addition, various tumor cells constitutively express G-CSF.

Human G-CSF receptor (G-CSF R) is a 120 kDa type I transmembrane glycoprotein that belongs to the haematopoietin receptor superfamily. The mature protein consists of a 603 amino acid extracellular domain (ECD), a 23 amino acid transmembrane fragment, and a 186 amino acid cytoplasmic domain. ECD contains an N-terminal Ig like domain, a cytokine receptor homology domain, and three fibronectin type III domains. Alternative splicing of human G-CSF r generates other isoforms, including potentially soluble forms of the receptor. The ECDs of mouse and human G-CSF r share 63% amino acid sequence identity. G-CSF r forms a complex with the ligand in a 2:2 ratio. It is expressed on monocytes, neutrophils, megakaryocytes, platelets, bone marrow progenitor cells, trophoblasts and placentas, endothelial cells, and various tumor cell types.

G-CSF is an important regulator of granulopoiesis in vivo, and mutations in G-CSF R are associated with congenital neutropenia. G-CSF can support the growth of multilineage hematopoietic progenitor cells and mobilize them from the bone marrow into the blood. G-CSF enhances the functional capacity of mature neutrophils and supports their survival by limiting the apoptosis rate. G-CSF also enhanced m-csf-induced monocyte generation caused by hematopoietic progenitor cells and stimulated the proliferation of peripheral Th2 inducing dendritic cells. It promotes the development of T cell immune tolerance and tissue recovery after myocardial infarction and cerebral ischemia. The human G-CSF immunoassay is a 3.5-4.5-hour solid-phase ELISA designed to measure human G-CSF in cell culture supernatants, serum, and plasma. It contains recombinant human G-CSF expressed in E. coli and antibodies against the recombinant protein. Recombinant human G-CSF has been shown to be accurately quantified. The results obtained using natural human G-CSF showed that the linear curve was parallel to the standard curve obtained using the kit standard. These results indicate that the kit can be used to determine the relative mass value of natural human G-CSF.