< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
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.
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
The recovery of TNF RII spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Mouse Cell Culture Supernates (n=5)
Mouse EDTA Plasma (n=5)
Mouse Serum (n=5)
Rat Cell Culture Supernates (n=5)
Rat EDTA Plasma (n=5)
Rat Serum (n=5)
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.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TNF RII/TNFRSF1B
TNF RII (Tumor Necrosis Factor Receptor II), also known as TNFRSF1B, p75/p80, and CD120b, is a widely expressed receptor for membrane-associated TNF-alpha and Lymphotoxin-alpha. Its activation initiates pro-inflammatory and pro-survival responses via NFkB-dependent signaling pathways, although it may also induce apoptosis.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
100 µL Conjugate
Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 5 times.
100 µL Substrate Solution
Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
100 µL Stop Solution
Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.