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TNF alpha ELISA Kit

TNF alpha Reactivity: Human Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Catalog No. ABIN6960272
  • Target See all TNF alpha ELISA Kits
    TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))
    Reactivity
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    • 12
    • 9
    • 5
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
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    Human
    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 human serum, plasma, tissue homogenates, cell lysates, cell culture supernates.
    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)
    Sensitivity
    6.5 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
    Top Product
    Discover our top product TNF alpha ELISA Kit
  • 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 1.0 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 0.5 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
  • Bassani, Bonato, Machado, Cóppola-Segovia, Moura, Zanata, Oliveira, Vital: "Decrease in Adult Neurogenesis and Neuroinflammation Are Involved in Spatial Memory Impairment in the Streptozotocin-Induced Model of Sporadic Alzheimer's Disease in Rats." in: Molecular neurobiology, Vol. 55, Issue 5, pp. 4280-4296, (2019) (PubMed).

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    Guo, Wang, Wang, Qiao, Tang: "Pantoprazole blocks the JAK2/STAT3 pathway to alleviate skeletal muscle wasting in cancer cachexia by inhibiting inflammatory response." in: Oncotarget, Vol. 8, Issue 24, pp. 39640-39648, (2018) (PubMed).

    Zhang, Hu, Sun, Li, Liu, Song: "Inhibitory Effect of Low-Intensity Pulsed Ultrasound on the Expression of Lipopolysaccharide-Induced Inflammatory Factors in U937 Cells." in: Journal of ultrasound in medicine : official journal of the American Institute of Ultrasound in Medicine, Vol. 36, Issue 12, pp. 2419-2429, (2018) (PubMed).

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    Bi, Sun, Wu, Chen, Tang, Mao: "PPARγ alleviated hepatocyte steatosis through reducing SOCS3 by inhibiting JAK2/STAT3 pathway." in: Biochemical and biophysical research communications, Vol. 498, Issue 4, pp. 1037-1044, (2018) (PubMed).

    Luo, Wang, Liu, Yin, A, Huang, Ye: "Inhibition of SOX9 Promotes Inflammatory and Immune Responses of Dental Pulp." in: Journal of endodontics, Vol. 44, Issue 5, pp. 792-799, (2018) (PubMed).

    Zhu, Liu, Zhang, Pan, Lin, Li, Wang, Yang, Yan, Gong: "Circulating and Adipose Tissue mRNA Levels of Zinc-α2-Glycoprotein, Leptin, High-Molecular-Weight Adiponectin, and Tumor Necrosis Factor-Alpha in Colorectal Cancer Patients With or Without Obesity." in: Frontiers in endocrinology, Vol. 9, pp. 190, (2018) (PubMed).

    Waldner, Dassati, Redl, Smania, Gandolfi: "Apolipoprotein D Concentration in Human Plasma during Aging and in Parkinson's Disease: A Cross-Sectional Study." in: Parkinson's disease, Vol. 2018, pp. 3751516, (2018) (PubMed).

    Gong, Guan, Tian, Li, Zhang: "Rho Kinase Type 1 (ROCK1) Promotes Lipopolysaccharide-induced Inflammation in Corneal Epithelial Cells by Activating Toll-Like Receptor 4 (TLR4)-Mediated Signaling." in: Medical science monitor : international medical journal of experimental and clinical research, Vol. 24, pp. 3514-3523, (2018) (PubMed).

    Zhou, He, Ping, Li, Zhu, Cui, Feng, Zhao, Zhang, Li, Sun: "Uncoupling Protein 2 and Peroxisome Proliferator-Activated Receptor γ Gene Polymorphisms in Association with Diabetes Susceptibility in Chinese Han Population with Variant Glucose Tolerance." in: International journal of endocrinology, Vol. 2018, pp. 4636783, (2018) (PubMed).

    Li, Ran, Zhang, Chen, Li, Luo, Wang, Jia: "Elevated serum milk fat globule-epidermal growth factor 8 levels in type 2 diabetic patients are suppressed by overweight or obese status." in: IUBMB life, Vol. 69, Issue 2, pp. 63-71, (2017) (PubMed).

    Mern, Tschugg, Hartmann, Thomé et al.: "Self-complementary adeno-associated virus serotype 6 mediated knockdown of ADAMTS4 induces long-term and effective enhancement of aggrecan in degenerative human nucleus pulposus cells: A new ..." in: PLoS ONE, Vol. 12, Issue 2, pp. e0172181, (2017) (PubMed).

    Abduljabbar, Vohra, Kellesarian, Javed et al.: "Efficacy of scaling and root planning with and without adjunct Nd:YAG laser therapy on clinical periodontal parameters and gingival crevicular fluid interleukin 1-beta and tumor necrosis factor-alpha ..." in: Journal of photochemistry and photobiology. B, Biology, Vol. 169, pp. 70-74, (2017) (PubMed).

  • Target See all TNF alpha ELISA Kits
    TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))
    Alternative Name
    Tumor Necrosis Factor Alpha (TNFa) (TNF alpha Products)
    Synonyms
    DIF ELISA Kit, TNF-alpha ELISA Kit, TNFA ELISA Kit, TNFSF2 ELISA Kit, RATTNF ELISA Kit, Tnfa ELISA Kit, tnf ELISA Kit, TNF-a ELISA Kit, TNFalpha ELISA Kit, Tnfsf1a ELISA Kit, TNFa ELISA Kit, cTNF ELISA Kit, Tnf-alpha ELISA Kit, tnfa-like ELISA Kit, TNF-ALPHA ELISA Kit, dif ELISA Kit, tnfa ELISA Kit, xtnf ELISA Kit, tnfsf2 ELISA Kit, tnf-alpha ELISA Kit, Cachectin ELISA Kit, tumor necrosis factor ELISA Kit, tumor necrosis factor b (TNF superfamily, member 2) ELISA Kit, tumor necrosis factor alpha ELISA Kit, tumor necrosis factor a (TNF superfamily, member 2) ELISA Kit, TNF ELISA Kit, Tnf ELISA Kit, tnf ELISA Kit, tnfb ELISA Kit, tnf-alpha ELISA Kit, LOC103694380 ELISA Kit, tnfa ELISA Kit
    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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