Key features and details
- One-wash 90 minute protocol
- Sensitivity: 1.04 pg/ml
- Range: 18.75 pg/ml - 1200 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Rat
Product nameRat TNF alpha ELISA Kit
See all TNF alpha kits
Intra-assay Sample n Mean SD CV% Supernatant 3 3.9% Inter-assay Sample n Mean SD CV% Supernatant 8 5.1%
Sample typeCell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma
Assay typeSandwich (quantitative)
Range18.75 pg/ml - 1200 pg/ml
Sample specific recovery Sample type Average % Range Cell culture supernatant 106 101% - 115% Serum 81 79% - 83% Hep Plasma 77 76% - 78% EDTA Plasma 81 81% - 82% Cit plasma 82 80% - 83%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Rat
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 (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
PlatformPre-coated microplate (12 x 8 well strips)
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10 x 96 tests 10X Rat TNF alpha Capture Antibody 1 x 600µl 1 x 6000µl 10X Rat TNF alpha Detector Antibody 1 x 600µl 1 x 6000µl 10X Wash Buffer PT (ab206977) 1 x 20ml 1 x 200ml Antibody Diluent 4BR 1 x 6ml 10 x 6ml Plate Seals 1 unit 1 x 10 units Rat TNF alpha Lyophilized Recombinant Protein 2 vials 2 x 10 vials Sample Diluent NS (ab193972) 1 x 50ml 2 x 250ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit 1 x 10 units Stop Solution 1 x 12ml 1 x 120ml TMB Development Solution 1 x 12ml 1 x 120ml
FunctionCytokine 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.
Involvement in diseaseGenetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis).
Sequence similaritiesBelongs to the tumor necrosis factor family.
modificationsThe soluble form derives from the membrane form by proteolytic processing.
The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.
Cellular localizationSecreted and Cell membrane.
- Information by UniProt
- APC1 protein
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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.
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.
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.
To learn more about the advantages of recombinant antibodies see here.
ab236712 has been referenced in 38 publications.
- Guo J et al. Puerarin attenuates preeclampsia-induced trophoblast mobility loss and inflammation by modulating miR-181b-5p/RBAK axis. Am J Reprod Immunol 87:e13510 (2022). PubMed: 34747081
- Zhang Y et al. Attenuation of hepatic ischemia‑reperfusion injury by adipose stem cell‑derived exosome treatment via ERK1/2 and GSK‑3β signaling pathways. Int J Mol Med 49:N/A (2022). PubMed: 34878156
- Huang X et al. Histone methyltransferase enhancer of zeste 2 polycomb repressive complex 2 subunit exacerbates inflammation in depression rats by modulating microglia polarization. Bioengineered 13:5509-5524 (2022). PubMed: 35172677
- Song J & Wang J SIRT3 regulates bronchial epithelium apoptosis and aggravates airway inflammation in asthma. Mol Med Rep 25:N/A (2022). PubMed: 35234263
- Park YJ et al. Effects of Glucocorticoid Therapy on Sepsis Depend Both on the Dose of Steroids and on the Severity and Phase of the Animal Sepsis Model. Life (Basel) 12:N/A (2022). PubMed: 35330172