Background:SCF/c-Kit Ligand
Stem cell factor (SCF), also known as c-kit ligand (KL), mast cell growth factor (MGF), and steel factor (SLF), is a widely expressed 28 - 40 kDa type I transmembrane glycoprotein. It promotes the survival, differentiation, and mobilization of multiple cell types including myeloid, erythroid, megakaryocytic, lymphoid, germ cell, and melanocyte progenitors. SCF is a primary growth and activation factor for mast cells and eosinophils. Mature human SCF consists of a 189 amino acid (aa) extracellular domain (ECD), a 23 aa transmembrane segment, and a 36 aa cytoplasmic tail. The ECD shows both N-linked and O-linked glycosylation. Proteolytic cleavage at two alternate sites in the extracellular juxtamembrane region releases a 25 kDa soluble molecule which is comparable to the only form produced by Steel-dickie mutant mice. An alternately spliced isoform of human SCF lacks 28 aa that encompasses the primary proteolytic recognition site. Within the ECD of the short isoform (corresponding to this recombinant protein), human SCF shares 75 - 83% aa sequence identity with canine, feline, mouse, and rat SCF. Rat SCF is active on mouse and human cells, but human SCF is only weakly active on mouse cells. Noncovalent dimers of transmembrane or soluble SCF interact with the receptor tyrosine kinase SCF-R/c-kit to trigger receptor dimerization and signaling. SCF assists in the recovery of cardiac function following myocardial infarction by increasing the number of cardiomyocytes and vascular channels.
原厂资料:
Product Summary:
The Quantikine Mouse SCF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse SCF levels in cell culture supernates, serum, and EDTA plasma. It contains E. coli-expressed recombinant mouse SCF and antibodies raised against the recombinant protein. Results obtained for naturally occurring mouse SCF showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values of natural mouse SCF.
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, EDTA Plasma
Intra-Assay Precision
Inter-Assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
20
20
20
Mean
67.2
183
1,038
73.4
193
1,033
Standard Deviation
2.5
6
30.5
5.3
8.9
47.4
CV%
3.7
3.3
2.9
7.2
4.6
4.6
Recovery:
The recovery of mouse SCF spiked to three levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Serum (n=5)
101
92-115
Cell Culture Supernates (n=5)
105
99-113
EDTA Plasma (n=4)
102
84-113
Linearity:
To assess the linearity of the assay, five or more samples spiked with various concentrations of mouse SCF in each matrix were diluted with Calibrator Diluent and then assayed.