Interleukin-1 (IL-1) is a name that designates two proteins, IL-1 alpha and IL-1 beta, which are the products of distinct genes, but which show approximately 25% amino acid sequence identity and which recognize the same cell surface receptors. Although IL-1 production is generally considered to be a consequence of inflammation, recent evidence suggests that IL-1 is also temporarily upregulated during bone formation and the menstrual cycle and can be induced in response to nervous system stimulation. In response to classic stimuli produced by inflammatory agents, infections or microbial endotoxins, a dramatic increase in the production of IL-1 by macrophages and various other cells is seen. Cells in particular known to produce IL-1 include osteoblasts, monocytes, macrophages, keratinocytes, Kupffer cells, hepatocytes, thymic and salivary gland epithelium, Schwann cells, fibroblasts and glia (oligodendroglia, astrocytes and microglia).
IL-1 alpha and IL-1 beta are both synthesized as 31 kDa precursors that are subsequently cleaved into proteins with molecular weights of approximately 17,000. Neither precursor contains a typical hydrophobic signal peptide sequence and most of the precursor form of IL-1 alpha remains in the cytosol of cells, although there is evidence for a membrane-bound form of the precursor form of IL-1 alpha. The IL-1 alpha precursor reportedly shows full biological activity in the EL 4 assay. Among various species, the amino acid sequence of mature IL-1 alpha is conserved 60% to 70% and human IL-1 has been found to be biologically active on murine cell lines. Both forms of IL-1 bind to the same receptors, designated type I and type II. Recent evidence suggests that only the type I receptor is capable of signal transduction and that the type II receptor may function as a decoy, binding IL-1 and thus preventing binding of IL-1 to the type I receptor.
原厂资料:
Product Summary:
The Quantikine Rat IL-1 alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure rat IL-1 alpha levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant rat IL-1 alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant rat IL-1 alpha accurately. Results obtained using natural rat IL-1 alpha showed dose 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 rat IL-1 alpha.
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
54
134
417
54
130
449
Standard Deviation
3.6
6.3
16.5
5
9.5
29.7
CV%
6.7
4.7
4
9.3
7.3
6.6
Recovery:
The recovery of rat IL-1 alpha spiked to three levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Serum (n=4)
92
85-99
Cell Culture Supernates (n=4)
104
100-111
EDTA Plasma (n=5)
98
89-112
Linearity:
To assess the linearity of the assay, samples spiked with rat IL-1 alpha in each matrix were diluted with Calibrator Diluent and then assayed.