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Your Position: Casa > Kits > TNF-alpha > RES-A027

resDetect™ Human Tumor Necrosis Factor Alpha (TNF-α) ELISA Kit (Residue Testing)

For research use only.

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Product Details
Assay TypeSandwich-ELISA
AnalyteTNF-α
Format96T
ReactivityHuman
Regulatory StatusRUO
Sensitivity< 6.25 pg/mL
Standard Curve Range6.25-400 pg/mL
Assay Time2 hr 50 min
Suitable Sample TypeFor the quantitative determination of human TNF-α in Cell Culture Supernatants, Plasma, Serum.
Sample volume100 uL
Materials Provided
IDComponentsSize
RES027-C01Pre-coated Anti-TNF-α Antibody Microplate1 plate
RES027-C02Human TNF-α Standard15 μg
RES027-C03Biotin-Anti-TNF-α Antibody20 μg
RES027-C04Streptavidin-HRP50 μL
RES027-C0510xWashing Buffer 50 mL
RES027-C062xDilution Buffer50 mL
RES027-C07Substrate Solution12 mL
RES027-C08Stop Solution7 mL
  • Background
    Tumour Necrosis Factor alpha (TNF-α) ELISA Kit is based on ELISA sandwich method and designed to measure human TNF-α levels in cell culture supernates, serum, and plasma. It contains recombinant human TNF-α and a pair of antibodies against the recombinant factor, which are provided by ACROBiosystems. Results are obtained by four parameter logistic curve that were parallel to the standard curves obtained. The verification results indicate that this kit can be used for the quantitative determination of GMP human Tumour Necrosis Factor alpha (TNF-α) (ACROBiosystems, cat#GMP-TNAH23) concentrations. The specificity has been verified.
  • Application

    The Human Tumor Necrosis Factor Alpha (TNF-α) (Residue Testing)was developed for the detection and quantitative determination of GMP human TNF-α in samples from CAR-T product preparation processing.

    It is for research use only.

  • Reconstitution
    Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • Storage
    Unopened kit should be stored at 2°C -8°C upon receiving.

    Find the expiration date on the outside packaging and do not use reagents past their expiration date.

    The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

  • Assay Principles
    This assay kit employs a standard sandwich-ELISA format, providing a rapid detection of Human TNF-α. The kit consists of Pre-coated Anti-TNF-α Antibody Microplate and Human TNF-α Standard and Biotin-Anti-TNF-α Antibody and Streptavidin-HRP and buffers.

    Your experiment will include 6 simple steps:

    a) Bring all reagents to room temperature(20℃-25℃) before use.

    b) Add your sample to the plate and take the Human TNF-α as standard. The samples and standard are diluted by Dilution Buffer.

    c) Wash the plate and add the Biotin-Anti-TNF-α Antibody diluted by Dilution Buffer to the plate.

    d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.

    e) Wash the plate and add TMB.

    f) Stop the substrate reaction by adding diluted acid. Absorbance (OD) is calculated by the absorbance at 450 nm minus the absorbance at 630 nm to remove background disturbance before statistical analysis. The OD Value reflects the amount of bound protein.

Typical Data Please refer to Ds document for the assay protocol.
 TNF-alpha TYPICAL DATA

For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.

Validation
Dilution Linearity

To assess the linearity of the assay, samples spiked with high concentrations of human TNF-α, were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

 TNF-alpha DILUTION LINEARITY
Intra-Assay Statistics

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision, Intra-Assay Precision CV<10%.

 TNF-alpha INTRA-ASSAY STATISTICS
Inter-Assay Statistics

Three samples of known concentration were tested in three separate assays to assess inter-assay precision, Inter-Assay Precision CV<10%.

 TNF-alpha INTER-ASSAY STATISTICS
Recovery

Three parts of blank serum were added with different concentrations of human TNF-α, and the serum without human TNF-α was used as background to calculate the recovery rate. The range of the recovery rate is 89.2-116.0%, and the average recovery is 98.3%.

 TNF-alpha RECOVERY
  • Background: TNF-alpha
    DIF,TNF-alpha,TNFA,TNFSF2,cachexin,cachectin,TNFα
    Tumor necrosis factor alpha (TNFα) is a cytokine produced primarily by monocytes and macrophages. It is found in synovial cells and macrophages in the tissues.The primary role of TNFα is in the regulation of immune cells. TNFα is able to induce apoptotic cell death, to induce inflammation, and to inhibit tumorigenesis and viral replication. Dysregulation of TNFα production has been implicated in a variety of human diseases, including major depression, Alzheimer's disease and cancer. Recombinant TNFα is used as an immunostimulant under the INN tasonermin. TNFα can be produced ectopically in the setting of malignancy and parallels parathyroid hormone both in causing secondary hypercalcemia and in the cancers with which excessive production is associated.
  • Clinical and Translational Updates

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