Background:G-CSF
G-CSF is a pleiotropic cytokine best known for its specific effects on the proliferation, differentiation, and activation of hematopoietic cells of the neutrophilic granulocyte lineage. It is produced mainly by monocytes and macrophages upon activation by endotoxin, TNF-alpha and IFN-gamma. Other cell types, including fibroblasts, endothelial cells, astrocytes and bone marrow stromal cells, can also secrete G-CSF after LPS, IL-1 or TNF-alpha activation. In addition, various carcinoma cell lines and myeloblastic leukemia cells can express G-CSF constitutively. In humans, two distinct cDNA clones for G-CSF, encoding 207 and 204 amino acid precursor proteins, have been isolated. Both proteins have a 30 amino acid signal peptide and have identical amino acid sequences except for a three amino acid insertion (deletion) at the 35th amino acid residue from the N-terminus of the mature protein. Human G-CSF is 73% identical at the amino acid level to mouse G-CSF and the two proteins show species cross-reactivity. In vitro, G-CSF stimulates growth, differentiation and functions of cells from the neutrophil lineage. It also has blast cell growth factor activity and can synergize with IL-3 to shorten the Go period of early hematopoietic progenitors. Consistent with itsin vitro functions, G-CSF has been found to play important roles in defense against infection, in inflammation and repair, and in the maintenance of steady state hematopoiesis.
原厂资料:
Product Summary:
The Quantikine Mouse G-CSF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse G-CSF in cell culture supernates and serum. It contains E. coli-expressed recombinant mouse G-CSF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse G-CSF accurately. Results obtained using natural mouse G-CSF showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse G-CSF.
Precision:
Intra-Assay Precision (Precision within an assay)Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays)Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum
Intra-Assay Precision
Inter-Assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
20
20
20
Mean
64.4
196
697
64.8
195
620
Standard Deviation
2.3
8.8
43
4.1
15.5
50.6
CV%
3.6
4.5
6.2
6.3
7.9
8.2
Recovery:
The recovery of mouse G-CSF spiked to three levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Serum (n=5)
96
80-119
Cell Culture Supernates (n=7)
92
82-110
Linearity:
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse G-CSF in each matrix were diluted with Calibrator Diluent and then assayed.