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Your Position: Home > Kits > TPO > RES-A034

resDetect™ Human TPO ELISA Kit (Residue Testing)

For research use only.

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    Product Details
    Assay TypeSandwich-ELISA
    AnalyteTPO
    Format96T
    ReactivityHuman
    Regulatory StatusRUO
    Sensitivity<12.5 pg/mL
    Standard Curve Range12.5 pg/mL-400 pg/mL
    Assay Time3 hr 20 min
    Suitable Sample TypeFor the detection and quantitative determination of TPO in human serum and cell culture supernates.
    Sample volume100 uL
  • Background
    Human TPO Kit is based on ELISA sandwich method and designed to measure human TPO levels in cell culture supernates, serum, and plasma. It contains recombinant human TPO and a pair of antibodies against the recombinant human TPO, 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 TPO (ACROBiosystems, cat#GMP-THNH25) concentrations. The specificity has been verified.
  • Application

    The kit is developed for  the detection and quantitative determination of TPO in human serum and cell culture supernates.

    It is for research use only.

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

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

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

    Materials Provided
    IDComponentsSize
    RES034-C01Pre-coated Anti-TPO Antibody Microplate1 plate
    RES034-C02Human TPO Standard20 μg
    RES034-C03Biotin-Anti-TPO Antibody20 μg
    RES034-C04Streptavidin-HRP50 μL
    RES034-C0510×Washing Buffer50 mL
    RES034-C062×Dilution Buffer50 mL
    RES034-C07Substrate Solution12 mL
    RES034-C08Stop Solution7 mL
  • Assay Principles
    This assay kit employs a standard sandwich-ELISA format, providing a rapid detection of Human TPO. The kit consists of Pre-coated Anti-TPO Antibody Microplate and Human TPO Standard and Biotin-Anti-TPO 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 TPO as standard. The samples and standard are diluted by Dilution Buffer.

    c) Wash the plate and add the Biotin-Anti-TPO 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.
 TPO 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 TPO were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

 TPO DILUTION LINEARITY
Intra-Assay Statistics

Three samples of known concentration were tested ten times on one plate to assess intra-assay precision.

 TPO INTRA-ASSAY STATISTICS
Inter-Assay Statistics

Three samples of known concentration were tested in three separate assays to assess inter-assay precision.

 TPO INTER-ASSAY STATISTICS
Recovery

Five parts of blank T cell culture supernatant were added with different concentrations of human TPO, and the T cell culture supernatant without human TPO was used as background to calculate the recovery rate. The range of the recovery rate is 94.7-108.9%, and the average recovery is 102.1%.

 TPO RECOVERY
  • Background: TPO
    Thrombopoietin (TPO) is a 332 amino acid glycoprotein made primarily in the liver and act as the major physiological regulator of platelet production. TPO binds the TPO receptor, activates JAK and STAT pathways, thus stimulating megakaryocyte growth and platelet production. Additionally, further investigation uncovered that thrombopoietin is a critical cytokine promoting hematopoietic rebound after myeloablation and its transcripts are expressed by multiple cellular sources.
  • Clinical and Translational Updates

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