Human sTNF RII/TNFRSF1B Quantikine ELISA Kit
Human sTNF RII/TNFRSF1B Quantikine ELISA Kit
Human sTNF RII/TNFRSF1B Quantikine ELISA Kit Summary
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货号：DRT200
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 uL), Serum (20 uL), EDTA Plasma (20 uL), Heparin Plasma (20 uL), Citrate Plasma (20 uL), Urine (20 uL)
Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
Specificity
Natural and recombinant human sTNF RII
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Product Summary
The Quantikine Human sTNF RII Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure human sTNF RII in cell culture supernates, serum, plasma, or urine. It contains E. coli-expressed recombinant human sTNF RII as well as antibodies raised against this polypeptide. The recombinant protein represents the non-glycosylated, N-terminal methionyl form of the naturally occurring human soluble Type II receptor for TNF, minus 53 amino acids from the proline-rich region of the protein just exterior to the transmembrane domain, with an apparent molecular weight of approximately 19.8 kDa. This immunoassay has been shown to accurately quantitate the recombinant soluble sTNF RII. Results obtained using natural sTNF 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 sTNF RII. Since the measurement of human sTNF RII by this immunoassay is insensitive to added TNF-alpha or TNF-beta, it is probable that this measurement corresponds to the total amount of the soluble receptor present in samples, (i.e. the total amount of free receptor plus the total amount of receptor bound to TNF).
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
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
|
|
Intra-Assay Precision
|
Inter-Assay Precision
|
|
Sample
|
1
|
2
|
3
|
1
|
2
|
3
|
|
n
|
20
|
20
|
20
|
20
|
20
|
20
|
|
Mean
|
68.7
|
179
|
349
|
89.8
|
197
|
444
|
|
Standard Deviation
|
2.2
|
4.6
|
16.8
|
4.6
|
6.9
|
16.2
|
|
CV%
|
3.2
|
2.6
|
4.8
|
5.1
|
3.5
|
3.6
|
Cell Culture Supernates, Urine
|
|
Intra-Assay Precision
|
Inter-Assay Precision
|
|
Sample
|
1
|
2
|
3
|
1
|
2
|
3
|
|
n
|
20
|
20
|
20
|
20
|
20
|
20
|
|
Mean
|
45.3
|
126
|
254
|
97.1
|
199
|
430
|
|
Standard Deviation
|
2
|
3.7
|
10.4
|
3.1
|
6.9
|
17.8
|
|
CV%
|
4.4
|
2.9
|
4.1
|
3.2
|
3.5
|
4.1
|
Recovery
The recovery of human sTNF RII spiked to levels throughout the range of the assay was evaluated.
|
Sample Type
|
Average % Recovery
|
Range %
|
|
Cell Culture Media (n=4)
|
96
|
88-103
|
|
Citrate Plasma (n=4)
|
100
|
91-116
|
|
EDTA Plasma (n=4)
|
95
|
85-100
|
|
Heparin Plasma (n=4)
|
89
|
74-108
|
|
Serum (n=4)
|
97
|
86-107
|
|
Urine (n=4)
|
97
|
83-111
|
