Human sTNF RII/TNFRSF1B Quantikine ELISA

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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)

Sensitivity

2.3 pg/mL

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

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