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Human TNF alpha ELISA Kit

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Referentie ELH-TNFa-1

Formaat : 1PlateKit

Merk : RayBiotech


Human TNF alpha ELISA Kit

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RayBio® Human TNF alpha ELISA Kit for cell culture supernatants, plasma, serum, and lysate samples. 30 pg/ml sensitivity.

Product Description

Specifications

Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Estimated Lead Time 1-2 business days
Species Human
Sensitivity 30 pg/ml

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Detection Range 30 pg/ml - 6000 pg/ml
Specificity This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human ANG, CD23, Eotaxin, GCSF, GM-CSF, GRO-alpha, GRO-beta, GRO-gamma, I-309, IFN-gamma, IL-1 alpha, IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-15, IL-16, IP-10, MCP-1, MCP-2, MCP-3, MCP-4, MCSF, MIG, MIP-1 alpha, MIP-1 beta, NAP-2, PDGF, PF-4.
Compatible Sample Types Cell Culture Supernatants, Plasma, Serum, Tissue Lysates, Cell Lysates
Recommended Dilution (Serum/Plasma) 2 fold
Accession Number P01375
Gene Id 7124
Gene Symbols TNF|TNFA|TNFSF2
Protein Name / Synonyms Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a) [Cleaved into: Tumor necrosis factor, membrane form (N-terminal fragment) (NTF) Intracellular domain 1 (ICD1) Intracellular domain 2 (ICD2) C-domain 1 C-domain 2 Tumor necrosis factor, soluble form]
Quantitative/Semi-Quantitative Quantitative
Solid Support 96-well Microplate
Design Principle Sandwich-based
Method Of Detection Colorimetric
Research Area Cardiovascular Disease, Cancer, Inflammation, Antibody Drug Development, Protein Therapeutics
Shipping Type Blue ice
Storage ≤-20°C

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    Target Background

    Tumor necrosis factor alpha (TNF alpha), also known as lymphotoxin and cachectin, is a pleiotropic cytokine. TNF alpha is a smaller cytokine with a molecular weight of 26 kDa (transmembrane) and 17 kDa (soluble) that binds to the same receptors producing a vast array of effects similar to those of TNF-beta. Both TNF-alpha and TNF-beta share 30% amino acid homology and have similar biological activities. TNF-alpha is produced primarily by monocytes, neutrophils and macrophages. It is found in synovial cells and macrophages in the tissues. TNF-alpha and TNF-beta can modulate many immune and inflammatory functions, while having the ability to inhibit tumor growth. Target tumor cells must express TNF receptors 1 (55 kDa) and 2 (75 kDa) to be killed, with the p55 receptor mediating the cytotoxic response.

    Product Features

    • Strip plates and additional reagents allow for use in multiple experiments
    • Quantitative protein detection
    • Establishes normal range
    • The best products for confirmation of antibody array data

    Standard Curves

    Standard curve graphs showing the results from the human tnf alpha elisa kit

    Dilution series diagram with 8 vials

    Spiking & Recovery Results

    Sample TypeAverage % RecoveryRange (%)
    Serum94.5382-103
    Plasma91.9280-102
    Cell culture media96.4384-104

    Linearity Results

      SerumPlasmaCell Culture Media
    1:2Average % of expected
    Range (%)    		94
    83-102    		93
    82-103    		92
    81-102
    1:4Average % of expected
    Range (%)    		94
    83-104    		92
    82-103    		99
    85-107

    Typical Data

    ELH-TNFa was used to measure plasma TNF levels in patients with axial spondyloarthritis and healthy controls. Figure adapted from Prokopcova et al., Heliyon, 2024.

    ELH-TNFa was used to measure TNF-α levels in lung tissue from immuno-humanized NCG mice following aerosol Mycobacterium tuberculosis infection. Figure adapted from Wang et al., EMBO Molecular Medicine, 2024.

    ELH-TNFa was used to measure soluble TNF levels in conditioned media from AZD5582-sensitive and -resistant oral squamous cell carcinoma cell lines. Figure adapted from Chai et al., Cancer Research Communications, 2024.

    ELH-TNFa-CL-1 was used to measure TNF-α levels in whole blood cell lysates from post-partum depression patients before and after brexanolone infusion. Figure adapted from Balan et al., eBioMedicine, 2023.

    ELH-TNFa was used to measure TNF-α secretion in culture supernatants from cytotoxic T lymphocytes activated by CD47 CAR-macrophages in a SKOV3 ovarian cancer model. Figure adapted from Chen et al., Journal of Translational Medicine, 2023.

    ELH-TNFa was used to measure TNF-α levels in culture supernatants from human umbilical vein endothelial cells exposed to silica nanoparticles. Figure adapted from Duan et al., International Journal of Nanomedicine, 2014.

    ELH-TNFa was used to measure TNF-α levels in serum-derived extracellular vesicles from colorectal cancer patients and healthy donors. Figure adapted from Xie et al., Archives of Biochemistry and Biophysics, 2023.

    Kit Components

    Additional Materials Required

    Assay Procedure Summary

    1. Prepare all reagents, samples and standards as instructed in the manual.
    2. Add 100 µL of standard or sample to each well.
    3. Incubate 2.5 h at RT or O/N at 4°C.
    4. Add 100 µL of prepared biotin antibody to each well.
    5. Incubate 1 h at RT.
    6. Add 100 µL of prepared Streptavidin solution to each well.
    7. Incubate 45 min at RT.
    8. Add 100 µL of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 µL of Stop Solution to each well.
    11. Read at 450 nm immediately.

    Storage/Stability

    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.

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