Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
31.2-2000 pg/mL
Sample Type
Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Mouse G-CSF DuoSet ELISA Features
- 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
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Background: G-CSF
Mouse
granulocyte-colony stimulating factor (G-CSF) is a 24-25 kDa monomeric
glycoprotein that regulates the proliferation, differentiation and activation
of hematopoietic cells. Mouse G-CSF cDNA encodes a 208 amino acid (aa)
precursor protein with a 30 aa signal sequence that is proteolytically cleaved
to form a 178 aa O-glycosylated mature protein containing two intrachain
disulfide bridges. In humans, two distinct cDNA clones, encoding a 204 aa form
and a minor alternatively spliced 207 aa form of G-CSF precursors, have been
isolated. Mouse and human G-CSF are 76% identical at the aa sequence level and
the two proteins show species cross-reactivity. G-CSF is produced primarily by
monocytes and macrophages upon activation by endotoxin, TNF-alpha or IL-1. Other
cell types, including fibroblasts, endothelial cells, astrocytes and bone
marrow stroma cells, can also secrete G-CSF after activation. In addition,
various tumor cells express G-CSF constitutively.
Mouse G-CSF
receptor (G-CSF R) is a 120 kDa, type I transmembrane glycoprotein that belongs
to the hematopoietin receptor superfamily. The mature protein is 812 amino
acids in length and contains a 601 aa extracellular region, a 24 aa
transmembrane segment, and a 187 aa cytoplasmic domain. The extracellular
region contains multiple modules, including an N-terminal Ig-like domain, a
cytokine receptor homology domain, and three fibronectin type III domains.
Based on crystallographic study, G-CSF receptor forms a complex with the ligand
in a 2:2 ratio. Mouse and human G-CSF receptor share 63% aa sequence identity.
Cells known to express G-CSF R include monocytes and neutrophils,
megakaryocytes and platelets, CD34+ CD33+ and CD34+ CD38+ hematopoietic
progenitors, trophoblasts, endothelial cells and various tumor cell types.
G-CSF is an
important regulator for granulopoiesis in vivo. It has been demonstrated that
G-CSF can support the growth of multi-lineage hematopoietic progenitor cells
without influencing their commitment to the myeloid lineage and mobilize
hematopoietic progenitor cells from the bone marrow into the bloodstream. On
mature neutrophils, G-CSF may regulate neutrophil survival by controlling their
rate of apoptosis. G-CSF has also been shown to enhance the functional capacity
of mature neutrophils. As a consequence of its effects on hematopoietic
progenitor cells, G-CSF has been shown to enhance monocytopoiesis in the
presence M-CSF. Within the peripheral blood stem cell population mobilized with
G-CSF, selective increases in the number of T helper 2-inducing dendritic cells
are found.
Long Name
Granulocyte Colony Stimulating Factor
Alternate Names
C17orf33, CSF3, CSF3OS, Filgrastim, GCSF, Lenograstim, Pluripoietin
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