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TNF alpha ELISA Kit (Tumor Necrosis Factor alpha) ELISA Kit

TNF alpha Reactivity: Rat Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Pubmed (138)
Catalog No. ABIN6574141
$560.00
Plus shipping costs $45.00
96 tests
local_shipping Shipping to: United States
Delivery in 9 to 11 Business Days
  • Target
    TNF alpha
    Reactivity
    • 17
    • 14
    • 11
    • 6
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Rat
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    15.6 pg/mL - 1000 pg/mL
    Minimum Detection Limit
    15.6 pg/mL
    Application
    ELISA
    Purpose
    The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of TNFa in rat serum, plasma, tissue homogenates, cell lysates, cell culture supernates.

    We offer validation data (WB) for the kit components. So you can be sure to order a reliable ELISA kit product composed of high quality reagents.
    Sample Type
    Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
    Analytical Method
    Quantitative
    Specificity
    This assay has high sensitivity and excellent specificity for detection of Tumor Necrosis Factor Alpha (TNFa)
    Cross-Reactivity (Details)
    No significant cross-reactivity or interference between Tumor Necrosis Factor Alpha (TNFa) and analogues was observed.
    Sensitivity
    5.9 pg/mL
    Components
    • Pre-coated, ready to use 96-well strip plate, flat buttom
    • Plate sealer for 96 wells
    • Reference Standard
    • Standard Diluent
    • Detection Reagent A
    • Detection Reagent B
    • Assay Diluent A
    • Assay Diluent B
    • Reagent Diluent (if Detection Reagent is lyophilized)
    • TMB Substrate
    • Stop Solution
    • Wash Buffer (30 x concentrate)
    • Instruction manual
  • Comment

    Information on standard material:
    The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

    Information on reagents:
    The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

    Information on antibodies:
    The provided antibodies and their host vary in different kits.

    Sample Volume
    100 μL
    Assay Time
    3 h
    Plate
    Pre-coated
    Protocol
    1. Prepare all reagents, samples and standards,
    2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
    3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
    4. Aspirate and wash 3 times,
    5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
    6. Aspirate and wash 5 times,
    7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
    8. Add 50μL Stop Solution. Read at 450nm immediately.
    Reagent Preparation
    1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
    2. Standard - Reconstitute the Standard with 0.5 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 1,000pg/mL. Prepare 7 tubes containing 0.25 mL Standard Diluent and produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 1,000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
    3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
    4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
    5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

    Note:

    1. Making serial dilution in the wells directly is not permitted.
    2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
    3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
    4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
    5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
    6. Contaminated water or container for reagent preparation will influence the detection result.
    Sample Preparation
    • It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
    • If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
    • If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
    • Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).
    Assay Precision
    Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
    CV(%) = SD/meanX100
    Intra-Assay: CV < 10%
    Inter-Assay: CV < 12%
    Restrictions
    For Research Use only
  • Precaution of Use
    The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
    Storage
    4 °C/-20 °C
    Storage Comment
    1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
    2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.
    .
    Expiry Date
    6 months
  • Ma, Zhao, Zhang, Chen: "Intra-arterial human urinary kallidinogenase alleviates brain injury in rats with permanent middle cerebral artery occlusion through PI3K/AKT/FoxO1 signaling pathway." in: Brain research, Vol. 1687, pp. 129-136, 2019 (PubMed).

    Yu, Wei, Weiss, Felder: "Angiotensin II Type 1a Receptors in the Subfornical Organ Modulate Neuroinflammation in the Hypothalamic Paraventricular Nucleus in Heart Failure Rats." in: Neuroscience, Vol. 381, pp. 46-58, 2019 (PubMed).

    Suluvoy, Sakthivel, Guruvayoorappan, Berlin Grace: "Protective effect of Averrhoa bilimbi L. fruit extract on ulcerative colitis in wistar rats via regulation of inflammatory mediators and cytokines." in: Biomedicine & pharmacotherapy, Vol. 91, pp. 1113-1121, 2018 (PubMed).

    Mohamed: "Protective Effect of Origanum Oil on Alterations of Some Trace Elements and Antioxidant Levels Induced by Mercuric Chloride in Male Rats." in: Biological trace element research, Vol. 182, Issue 1, pp. 49-56, 2018 (PubMed).

    Meng, Li, Li, He, Cui, Yin, Zhang, Li, Sang, Wang, Niu, Zhang, Guo, Sun, Wang, Bai, Xiao: "Cis-stilbene glucoside in Polygonum multiflorum induces immunological idiosyncratic hepatotoxicity in LPS-treated rats by suppressing PPAR-γ." in: Acta pharmacologica Sinica, Vol. 38, Issue 10, pp. 1340-1352, 2018 (PubMed).

    Xiao, Ye, Wang, Xiao, Fu, Xia, Zhang, Liu, Zeng: "Mild hypothermia pretreatment protects against liver ischemia reperfusion injury via the PI3K/AKT/FOXO3a pathway." in: Molecular medicine reports, Vol. 16, Issue 5, pp. 7520-7526, 2018 (PubMed).

    Li, Wu, Liu, Zhang, Bai, Chen: "The effect of anagliptin on intimal hyperplasia of rat carotid artery after balloon injury." in: Molecular medicine reports, Vol. 16, Issue 6, pp. 8003-8010, 2018 (PubMed).

    Li, Yang, Zhang, Liu, Zhao, Luo, Chen, Huang: "Ginsenoside Rg1 inhibits inflammatory responses via modulation of the nuclear factor‑κB pathway and inhibition of inflammasome activation in alcoholic hepatitis." in: International journal of molecular medicine, Vol. 41, Issue 2, pp. 899-907, 2018 (PubMed).

    Tan, Lin, Wang, Yang, Ma: "Cardioprotective time-window of Penehyclidine hydrochloride postconditioning: A rat study." in: European journal of pharmacology, Vol. 812, pp. 48-56, 2018 (PubMed).

    Zeng, Qiao, Lv, Fan, Liu, Xie: "High-dose dexamethasone induced LPS-stimulated rat alveolar macrophages apoptosis." in: Drug design, development and therapy, Vol. 11, pp. 3097-3104, 2018 (PubMed).

    Zhang, Xiong, Chen, Chen, Zhu, Li: "Therapeutic effect of bone marrow mesenchymal stem cells pretreated with acetylsalicylic acid on experimental periodontitis in rats." in: International immunopharmacology, Vol. 54, pp. 320-328, 2018 (PubMed).

    Song, Liu, Diao, Sun, Gao, Zhang, Chen, Pei: "Hydrogen‑rich solution against myocardial injury and aquaporin expression via the PI3K/Akt signaling pathway during cardiopulmonary bypass in rats." in: Molecular medicine reports, Vol. 18, Issue 2, pp. 1925-1938, 2018 (PubMed).

    Xiong, Ouyang, Jiang, Yang, Hu, Chen, Wang, Liu, Wang: "Chemical composition of Cyclocarya paliurus polysaccharide and inflammatory effects in lipopolysaccharide-stimulated RAW264.7 macrophage." in: International journal of biological macromolecules, Vol. 107, Issue Pt B, pp. 1898-1907, 2018 (PubMed).

    Elkhoely, Kamel: "Diallyl sulfide alleviates cisplatin-induced nephrotoxicity in rats via suppressing NF-κB downstream inflammatory proteins and p53/Puma signalling pathway." in: Clinical and experimental pharmacology & physiology, Vol. 45, Issue 6, pp. 591-601, 2018 (PubMed).

    Tian, Yang, Liu, Xu: "Emodin Attenuates Bleomycin-Induced Pulmonary Fibrosis via Anti-Inflammatory and Anti-Oxidative Activities in Rats." in: Medical science monitor : international medical journal of experimental and clinical research, Vol. 24, pp. 1-10, 2018 (PubMed).

    Said, Zaitone, Eldosoky, Elsherbiny: "Nifuroxazide, a STAT3 inhibitor, mitigates inflammatory burden and protects against diabetes-induced nephropathy in rats." in: Chemico-biological interactions, Vol. 281, pp. 111-120, 2018 (PubMed).

    Liu, Yang, Song, Geng, Chen: "Protective effects of N(2)‑L‑alanyl‑L‑glutamine mediated by the JAK2/STAT3 signaling pathway on myocardial ischemia reperfusion." in: Molecular medicine reports, Vol. 17, Issue 4, pp. 5102-5108, 2018 (PubMed).

    Zuo, Zhang, Mu, Shen, Li, Wang, Li, Chen: "Resolvin D2 protects against cerebral ischemia/reperfusion injury in rats." in: Molecular brain, Vol. 11, Issue 1, pp. 9, 2018 (PubMed).

    Shahin, Abdelkader, Safar: "A Novel Role of Irbesartan in Gastroprotection against Indomethacin-Induced Gastric Injury in Rats: Targeting DDAH/ADMA and EGFR/ERK Signaling." in: Scientific reports, Vol. 8, Issue 1, pp. 4280, 2018 (PubMed).

    Dong, Guo, Ji, Cao, Zhang, Chen, Huang, Wu, Lu, Sun: "S1PR3 is essential for phosphorylated fingolimod to protect astrocytes against oxygen-glucose deprivation-induced neuroinflammation via inhibiting TLR2/4-NFκB signalling." in: Journal of cellular and molecular medicine, Vol. 22, Issue 6, pp. 3159-3166, 2018 (PubMed).

  • Target
    TNF alpha
    Alternative Name
    Tumor Necrosis Factor Alpha (TNFa) (TNF alpha ELISA Kit Abstract)
    Synonyms
    DIF, TNF-alpha, TNFA, TNFSF2, RATTNF, Tnfa, tnf, TNF-a, TNFalpha, Tnfsf1a, TNFa, cTNF, Tnf-alpha, tnfa-like, TNF-ALPHA, dif, tnfa, xtnf, tnfsf2, tnf-alpha, Cachectin, tumor necrosis factor, tumor necrosis factor b (TNF superfamily, member 2), tumor necrosis factor alpha, tumor necrosis factor a (TNF superfamily, member 2), TNF, Tnf, tnf, tnfb, tnf-alpha, LOC103694380, tnfa
    UniProt
    P16599
    Pathways
    NF-kappaB Signaling, Apoptosis, Caspase Cascade in Apoptosis, TLR Signaling, 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, Inflammasome
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