Human G-CSF ARExSet ELISA

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 neutrophilic granulocyte lineage. Mature human G-CSF is a 178 amino acid (aa) O-glycosylated protein that contains two intrachain disulfide bridges. In humans, alternate splicing generates a second minor isoform with a 3 aa deletion. Mouse and human G-CSF share 76% aa 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 stroma cells. In addition, various tumor cells express G-CSF constitutively. Human G-CSF receptor (G-CSF R) is a 120 kDa type I transmembrane glycoprotein that belongs to the hematopoietin receptor superfamily. The mature protein consists of a 603 aa extracellular domain (ECD), a 23 aa transmembrane segment, and a 186 aa cytoplasmic domain. The ECD contains an N-terminal Ig-like domain, a cytokine receptor homology domain, and three fibronectin type III domains. Alternate splicing of human G-CSF R generates additional isoforms including a potentially soluble form of the receptor. The ECDs of mouse and human G-CSF R share 63% aa sequence identity. G-CSF R forms a complex with the ligand in a 2:2 ratio. It is expressed on monocytes, neutrophils, megakaryocytes, platelets, myeloid progenitors, trophoblasts and placenta, endothelial cells, and various tumor cell types. G-CSF is an important regulator for 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 bloodstream. G-CSF enhances the functional capacity of mature neutrophils and supports their survival by limiting the rate of apoptosis. G-CSF also enhances M-CSF induced monocytopoiesis from hematopoietic progenitor cells and stimulates the proliferation of peripheral Th2-inducing dendritic cells. It promotes the development of T cell immune tolerance as well as tissue recovery following myocardial infarction and cerebral ischemia.

pg/ml	O.D.		Average	Corrected
0.00	0.0219	0.0214	0.0217
31.25	0.1501	0.1242	0.1372	0.1155
62.50	0.2717	0.2517	0.2617	0.2401
125.00	0.4979	0.4999	0.4989	0.4773
250.00	0.9518	0.9266	0.9392	0.9176
500.00	1.8560	1.7680	1.8120	1.7904
1000.00	3.1680	2.9560	3.0620	3.0404
2000.00	4.2224	4.1105	4.1665	4.1448

• Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
• Development protocols are provided to guide further assay optimization
• Assay can be customized to your specific needs
• Economical alternative to complete kits

• Capture Antibody
• Detection Antibody
• Recombinant Standard
• Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

ARExSet^®Ancillary Reagent Kit (5 plates): containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and assay buffer.

96 well microplates: AREX Biosciences, Catalog # DSEP01. 

Plate Sealers: AREX Biosciences, Catalog # DSSF01.

Coating Buffer: 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2-7.4, 0.2 μm filtered . AREX Biosciences, Catalog # DSCB01.

Blocking Buffer: AREX Biosciences, Catalog # DSBB01.

Wash Buffer: 0.05% Tween 20 in PBS5, pH 7.2-7.4. AREX Biosciences, Catalog # DSWB01.

Assay Buffer: 0.5%BSA, 0.05%Tween20, PBS Solution.AREX Biosciences, Catalog # DSAB01

Substrate Solution: Tetramethylbenzidine. AREX Biosciences, Catalog # DSTS01.

Stop Solution: 0.5 mol/ml H₂SO₄. AREX Biosciences, Catalog # DSSS01.