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|Antigen||Tumor Necrosis Factor (TNF) ELISA Kits|
|Reactivity||Rat (Rattus) Alternatives|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||25-20000 pg/mL|
|Minimum Detection Limit||25 pg/mL|
|20 references available|
|Supplier||Log in to see|
Product Details Tumor Necrosis Factor ELISA KitTarget details Application Details Handling References for Tumor Necrosis Factor Kit (ABIN625216) Images
|Purpose||Rat TNF alpha ELISA Kit for cell culture supernatants, plasma, and serum samples.|
|Sample Type||Plasma, Cell Culture Supernatant, Serum|
|Specificity||This ELISA kit shows no cross-reactivity with any of the cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3 alpha, beta- NGF, TIMP-1, VEGF.|
|Cross-Reactivity (Details)||This ELISA kit shows no cross-reactivity with any of the cytokines tested (e.g., rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1alpha, IL- 1beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3alpha, beta- NGF, TIMP-1, VEGF).|
|Sensitivity||< 25 pg/mL|
|Material not included||
Target detailsProduct Details Tumor Necrosis Factor ELISA Kit Application Details Handling References for Tumor Necrosis Factor Kit (ABIN625216) Images back to top
|Alternative Name||TNF-alpha (TNF ELISA Kit Abstract)|
|Background||Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a)|
|Gene ID||103694380, 24835|
|Research Area||Virology, Atherosclerosis, Immunology, Receptors, Cytokines, Growth Factors, Innate Immunity, Cancer, Cardiovascular, Inflammation|
|Pathways||NF-kappaB Signaling, Apoptosis, Caspase Cascade in Apoptosis, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus|
Application DetailsProduct Details Tumor Necrosis Factor ELISA Kit Target details Handling References for Tumor Necrosis Factor Kit (ABIN625216) Images back to top
|Application Notes||Recommended Dilution for serum and plasma samples2 fold|
|Sample Volume||100 μL|
1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should beused for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 2 fold*. * Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
3. Assay Diluent B should be diluted 5-fold with deionized or distilled water.
4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 100 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 100 µL TNF-alpha standard from the vial of Item C, into a tube with 400 µL Assay Diluent A or 1x Assay Diluent B to prepare a 20,000 pg/mL stock standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 100 µL standard + 400 µL 200myl 200 µL 200 µL 200 µL 20,000 6,667 2,222 740.7 246.9 82.30 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 200-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 50 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent B to prepare a 200-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
5. Discard the solution. Repeat the wash as in step
6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
7. Discard the solution. Repeat the wash as in step
8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
|Calculation of Results||
Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Rat TNF-alpha concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 10 100 1,000 10,000 100,000 Assay Diluent B Rat TNF-alpha concentration (pg/mL) O D =4 50 n m 0.1 1 10 10 100 1,000 10,000 100,000
Sensitivity: The minimum detectable dose of TNF-alpha is typically less than 25 pg/mL.
Recovery: Recovery was determined by spiking various levels of Rat TNF-alpha into Rat serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 95.43 84-104 Plasma 93.79 82-102 Cell culture media 101.54 89-108
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93 94 98 Range ( %) 82-102 84-103 88-106 1:4 Average % of Expected 96 95 101 Range ( %) 85-104 83-103 90-108
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %
|Assay Precision||Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %|
|Restrictions||For Research Use only|
HandlingProduct Details Tumor Necrosis Factor ELISA Kit Target details Application Details References for Tumor Necrosis Factor Kit (ABIN625216) Images back to top
|Handling Advice||Avoid repeated freeze-thaw cycles.|
|Storage Comment||The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.|
|Expiry Date||6 months|
References for Tumor Necrosis Factor Kit (ABIN625216)Product Details Tumor Necrosis Factor ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
Xu, Yang, Sun, Chen, Jiang, Zheng, Ding, Liu, Sheng, Cui, Duan: "2,3',4,4',5-Pentachlorobiphenyl Induces Inflammatory Responses in the Thyroid Through JNK and Aryl Hydrocarbon Receptor-Mediated Pathway." in: Toxicological sciences : an official journal of the Society of Toxicology, Vol. 149, Issue 2, pp. 300-11, 2016
Rana, Sharma et al.: "Mechanism of sphingosine-1-phosphate induced cardioprotection against I/R injury in diabetic rat heart: Possible involvement of glycogen synthase kinase 3β and mitochondrial permeability transition ..." in: Clinical and experimental pharmacology & physiology, Vol. 43, Issue 2, pp. 166-73, 2016
Begum, Sharma, Pillai, Aeri, Sheliya: "Inhibitory effect of Careya arborea on inflammatory biomarkers in carrageenan-induced inflammation." in: Pharmaceutical biology, Vol. 53, Issue 3, pp. 437-45, 2015
Abdelkader, Safar, Salem: "Ursodeoxycholic Acid Ameliorates Apoptotic Cascade in the Rotenone Model of Parkinson's Disease: Modulation of Mitochondrial Perturbations." in: Molecular neurobiology, 2015
Sharma, Deshmukh: "Vinpocetine attenuates MPTP-induced motor deficit and biochemical abnormalities in Wistar rats." in: Neuroscience, Vol. 286, pp. 393-403, 2015
Khan, Jamwal, Bijjem, Prakash, Kumar: "Neuroprotective effect of hemeoxygenase-1/glycogen synthase kinase-3? modulators in 3-nitropropionic acid-induced neurotoxicity in rats." in: Neuroscience, Vol. 287C, pp. 66-77, 2015
Zhang, Zhang, Tsui: "Mesenteric lymph duct drainage attenuates acute lung injury in rats with severe intraperitoneal infection." in: Inflammation, Vol. 38, Issue 3, pp. 1239-49, 2015
Kabel, Abd Elmaaboud, Albarraq: "Ameliorative potential of omega 3 fatty acids and HMG-CoA reductase inhibitors on experimentally-induced non-alcoholic steatohepatitis." in: Prostaglandins, leukotrienes, and essential fatty acids, Vol. 96, pp. 1-9, 2015
Elshazly: "Ameliorative effect of nicorandil on high fat diet induced non-alcoholic fatty liver disease in rats." in: European journal of pharmacology, Vol. 748, pp. 123-32, 2015
Tyagi, Bisht, Pant, Kumar, Majeed, Prakash: "Possible role of GABA-B receptor modulation in MPTP induced Parkinson's disease in rats." in: Experimental and toxicologic pathology : official journal of the Gesellschaft für Toxikologische Pathologie, Vol. 67, Issue 2, pp. 211-7, 2015
Yaidikar, Thakur: "Punicalagin attenuated cerebral ischemia-reperfusion insult via inhibition of proinflammatory cytokines, up-regulation of Bcl-2, down-regulation of Bax, and caspase-3." in: Molecular and cellular biochemistry, Vol. 402, Issue 1-2, pp. 141-8, 2015
Aral, Kesim, Greenwell, Kara, Çetin, Yakan: "Alveolar bone protective and hypoglycemic effects of systemic propolis treatment in experimental periodontitis and diabetes mellitus." in: Journal of medicinal food, Vol. 18, Issue 2, pp. 195-201, 2015
Nassar, Al-Shorbagy, Arab, Abdallah: "Saxagliptin: a novel antiparkinsonian approach." in: Neuropharmacology, Vol. 89, pp. 308-17, 2015
Sayd, Antón, Alén, Caso, Pavón, Leza, Rodríguez de Fonseca, García-Bueno, Orio: "Systemic administration of oleoylethanolamide protects from neuroinflammation and anhedonia induced by LPS in rats." in: The international journal of neuropsychopharmacology / official scientific journal of the Collegium Internationale Neuropsychopharmacologicum (CINP), Vol. 18, Issue 6, 2015
Serya, Abbas, Ismail, Esmat, Abou El Ella: "Design, synthesis and biological evaluation of novel quinazoline-based anti-inflammatory agents acting as PDE4B inhibitors." in: Chemical & pharmaceutical bulletin, Vol. 63, Issue 2, pp. 102-16, 2015
Abdel Kawy: "Cilostazol attenuates cholestatic liver injury and its complications in common bile duct ligated rats." in: European journal of pharmacology, Vol. 752, pp. 8-17, 2015
Choudhary, Sheela Devi: "Longer period of oral administration of aspartame on cytokine response in Wistar albino rats." in: Endocrinologi?a y nutricio?n : o?rgano de la Sociedad Espan?ola de Endocrinologi?a y Nutricio?n, Vol. 62, Issue 3, pp. 114-22, 2015
Chen, Min, Wang, Leung, Shi, Zhou, Yu, Wang, An, Sha, Chen: "Pre-activation of mesenchymal stem cells with TNF-?, IL-1? and nitric oxide enhances its paracrine effects on radiation-induced intestinal injury." in: Scientific reports, Vol. 5, pp. 8718, 2015
Abdelsalam, Safar: "Neuroprotective effects of vildagliptin in rat rotenone Parkinson's disease model: role of RAGE-NF?B and Nrf2-antioxidant signaling pathways." in: Journal of neurochemistry, Vol. 133, Issue 5, pp. 700-7, 2015
Shete, Pawar, Thanki, Jain, Bansal: "Oral bioavailability and pharmacodynamic activity of hesperetin nanocrystals generated using a novel bottom-up technology." in: Molecular pharmaceutics, Vol. 12, Issue 4, pp. 1158-70, 2015
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