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.
Human TNF RII/TNFRSF1B Quantikine ELISA Kit
R&D Systems | Catalog # DRT200
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Key Product Details
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 µL), Serum (20 µL), EDTA Plasma (20 µL), Heparin Plasma (20 µL), Citrate Plasma (20 µL), Urine (20 µL)
Sensitivity
2.3 pg/mL
Assay Range
7.8-500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
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Product Summary for Human TNF RII/TNFRSF1B Quantikine ELISA Kit
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).
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
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.
Sample Values
Serum/Plasma/Urine - Samples from apparently healthy volunteers were evaluated for the
presence of human TNF RII in this assay. No medical histories were available for the donors
used in this study.
*The urine values are actual and are not normalized for creatinine content.
| Sample Type | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=40) | 1726 | 1003-3170 | 387 |
| EDTA plasma (n=40) | 1500 | 829-2262 | 320 |
| Heparin plasma (n=40) | 1678 | 951-2503 | 349 |
| Citrate plasma (n=40) | 1405 | 787-2145 | 286 |
| Urine* (n=27) | 1724 | 275-6982 | 1274 |
Cell Culture Supernates - Human peripheral blood mononuclear cells (1 x 106
cells/mL) were
cultured in RPMI supplemented with 10% fetal bovine serum, 50 μM beta -mercaptoethanol,
2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate and stimulated
with 10 μg/mL of PHA. Aliquots were removed on days 1, 3 and 5 and assayed.
| Day 1 (pg/mL) | Day 3 (pg/mL) | Day 5 (pg/mL) |
| 1841 | 2861 | 3124 |
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, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 45.3 | 126 | 254 | 97.1 | 199 | 430 |
| Standard Deviation | 2.0 | 3.7 | 10.4 | 3.1 | 6.9 | 17.8 |
| CV% | 4.4 | 2.9 | 4.1 | 3.2 | 3.5 | 4.1 |
Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 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 |
Recovery for Human TNF RII/TNFRSF1B Quantikine ELISA Kit
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 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of sTNF RII were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human TNF RII/TNFRSF1B Quantikine ELISA Kit
Human TNF RII/TNFRSF1B ELISA Cell Culture Supernate/Urine Standard Curve
Human TNF RII/TNFRSF1B ELISA Serum/Plasma Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TNF RII/TNFRSF1B
Long Name
Tumor Necrosis Factor Receptor II
Alternate Names
CD120b, TNFRII, TNFRSF1B
Gene Symbol
TNFRSF1B
Additional TNF RII/TNFRSF1B Products
Product Documents for Human TNF RII/TNFRSF1B Quantikine ELISA Kit
Product Specific Notices for Human TNF RII/TNFRSF1B Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human TNF RII/TNFRSF1B Quantikine ELISA Kit
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Sample Tested: UrineVerified Customer | Posted 11/10/2023Urine showed good linearity up to 160-fold dilution.
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Sample Tested: EDTA PlasmaVerified Customer | Posted 10/13/2021
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Sample Tested: PlasmaVerified Customer | Posted 02/03/2021Kit performed as intended with very clear protocol.
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Sample Tested: Jurkat human acute T cell leukemia cell lineVerified Customer | Posted 12/06/2019
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Protocols
View specific protocols for Human TNF RII/TNFRSF1B Quantikine ELISA Kit (DRT200):
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.
- Add 50 µL of Assay Diluent to each well.
- Add 200 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 twice for a total of 3 washes.
- Add 200 µL of Conjugate to each well.
- For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
For Cell Culture Supernate Samples & Urine Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour. - Aspirate and wash 3 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
200 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
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Associated Pathways
IL-15 Signaling Pathways and their Primary Biological Effects in Different Immune Cell Types
mTOR Signaling Pathway
TNF Superfamily Pathway: Human Ligand-Receptor Interactions & their Associated Functions
