Tumor Necrosis Factor Receptor-I (TNF RI, also known as p55/p60) and TNF RII (also known as p75/p80) are both high affinity receptors for TNF-alpha and -beta, potent mediators of multiple aspects of inflammatory immune responses. Both RI and RII are prototypic members of the TNF receptor superfamily and have been designated TNFRSF1A and TNFRSF1B, respectively. Whereas all cell types are thought to express TNF RI, TNF RII expression is limited primarily to hematopoietic cells and cells of the immune system. Most of the biological functions of TNF are mediated via TNF RI. Soluble forms of both TNF RI and TNF RII, generated as a result of proteolytic cleavage of the extracellular domains, have been detected in vivo in various biological fluids. The soluble TNF receptors can bind TNF with high affinity and functions as TNF antagonists.
TNF receptors are thought to form homotrimers or oligomers upon ligand binding. The monomers of both TNF receptors subtypes have similar extracellular domains including four cysteine-rich repeats characteristic of the TNF receptor superfamily. However, the intracellular domains differ, and consequently, the receptors exhibit different signaling capabilities. For instance, although both are capable of interacting with adaptor proteins upstream of NF-kB activation, only TNF RI has a cytoplasmic "death domain" upstream of caspase-dependent apoptosis cascades. Soluble forms of both TNF receptors can be generated via proteolytic cleavage.
原厂资料:
Product Summary:
The Quantikine Mouse/Rat TNF RII Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse or rat TNF RII in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse TNF RII and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant protein accurately. Results obtained using natural mouse or rat TNF RII 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 for natural mouse or rat TNF RII.
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
37.2
112
472
38.6
119
500
Standard Deviation
1.5
4.4
21
1.4
3.5
14.7
CV%
4
3.9
4.4
3.6
2.9
2.9
Recovery:
The recovery of TNF RII spiked to levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Mouse Cell Culture Supernates (n=5)
102
83-119
Rat Cell Culture Supernates (n=5)
98
85-116
Mouse Serum (n=5)
99
91-105
Rat Serum (n=5)
97
84-114
Mouse EDTA Plasma (n=5)
95
80-119
Rat EDTA Plasma (n=5)
95
83-112
Linearity:
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of TNF RII were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.