Rat TNF alpha ELISA Kit is a single-wash 90-min Simplestep used to quantify Rat TNF alpha with a sensitivity of 1.04 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
- Cited in over 60 citations
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation (By similarity). Induces insulin resistance in adipocytes via inhibition of insulin-induced IRS1 tyrosine phosphorylation and insulin-induced glucose uptake. Induces GKAP42 protein degradation in adipocytes which is partially responsible for TNF-induced insulin resistance (By similarity). Plays a role in angiogenesis by inducing VEGF production synergistically with IL1B and IL6 (By similarity). Promotes osteoclastogenesis and therefore mediates bone resorption (By similarity). The TNF intracellular domain (ICD) form induces IL12 production in dendritic cells.
Tnfa, Tnfsf2, Tnf, Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a
Rat TNF alpha ELISA Kit is a single-wash 90-min Simplestep used to quantify Rat TNF alpha with a sensitivity of 1.04 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
- Cited in over 60 citations
Sample | n | C.V. |
---|---|---|
Sample Supernatant | n 3 | C.V. 3.9 |
Sample | n | C.V. |
---|---|---|
Sample Supernatant | n 8 | C.V. 5.1 |
Sample type | Average % | Range |
---|---|---|
Sample type Cell culture supernatant | Average % = 106 | Range 101 - 115 % |
Sample type Serum | Average % = 81 | Range 79 - 83 % |
Sample type EDTA Plasma | Average % = 81 | Range 81 - 82 % |
Sample type Heparin Plasma | Average % = 77 | Range 76 - 78 % |
Sample type Citrate plasma | Average % = 82 | Range 80 - 83 % |
Rat TNF alpha ELISA Kit (ab236712) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of TNF alpha protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Rat TNF alpha with 1.04 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Example of rat TNF alpha standard curve in Sample Diluent NS.
The TNF alpha standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Interpolated concentrations of native TNF alpha in rat cell culture supernatant samples.
The concentrations of TNF alpha were measured in duplicates, interpolated from the TNF alpha standard curves and corrected for sample dilution. Undiluted samples are 50 ng/mL PMA + 500 ng/mL calcium ionomycin treated splenocyte cell culture supernatant - 100%. Treatment length was 72 hours. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was determined to be 156.05 pg/mL in treated splenocyte culture supernatant.
Interpolated concentrations of native TNF alpha in rat splenocyte cell culture supernatant based on 100% load.
The concentrations of TNF alpha were measured in duplicate and interpolated from the TNF alpha standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was below the lowest standard (<18.75) pg/mL in unstimulated splenocyte cell culture supernatant and 156.05 pg/mL in stimulated splenocyte cell culture supernatant. Stimulated samples were treated with 50 ng/mL PMA + 500 ng/mL calcium ionomycin for 72 hours. Unstimulated cells as a control were cultured in parallel for 72 hours.
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