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Your Position: 홈 > Kits > CD3 > RES-A095

resDetect™ Anti-CD3 Antibody Indirect ELISA Kit (Residue Testing)

For research use only.
    Materials Provided
    IDComponentsSize
    RES095-C01Pre-coated Human CD3E & CD3G Microplate1 plate
    RES095-C02Anti-CD3 Antibody Standard20 μg
    RES095-C03HRP-conjugated antibody10 μg
    RES095-C0410×Washing Buffer50 mL
    RES095-C052×Dilution Buffer50 mL
    RES095-C06Substrate Solution12 mL
    RES095-C07Stop Solution7 mL
  • Background
    Since the 1990s, CD3 monoclonal antibody has been used in CIK cell therapy to stimulates the proliferation and activation of T cells. Under the cooperation of other cytokines, such as IL2 and IL1a, CIK cells with rapid proliferation, high tumoricidal activity, broad tumor killing spectrum and non-MHC-restricted tumor killing characteristics are generated, which has significant effects on the treatment of cancer, chronic leukemia, liver disease and neurological diseases. Obviously, It is necessary to control the residues of raw materials in the final cell therapy product.
  • Application

    The kit is developed for the detection of anti-CD3 antibody in Bioprocess manufacturing applications. It is used as a tool to aid in routine quality control of in-process streams as well as final product .

    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.

  • Assay Principles
    This assay kit employs a standard indirect-ELISA format, providing a rapid detection of Anti-CD3 antibodies. The kit consists of Pre-coated Human CD3E & CD3G Microplate and Anti-CD3 Antibody Standard and HRP-conjugated antibody and related buffer.

    Your experiment will include 5 simple steps:

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

    b) Add your sample to the plate and take the Anti-CD3 Antibody as standard. The samples and standard are diluted by Dilution Buffer.

    c) Wash the plate and add the HRP-conjugated antibody diluted by Dilution Buffer to the plate.

    d) Wash the plate and add TMB.

    e) Stop the substrate reaction by adding diluted acid. Absorbance (OD) is calculated as 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 Antibody bound.

Typical Data Please refer to DS document for the assay protocol.
CD3 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 Anti-CD3 Antibody were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

CD3 DILUTION LINEARITY
Intra-Assay Statistics

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

CD3 INTRA-ASSAY STATISTICS
Inter-Assay Statistics

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

CD3 INTER-ASSAY STATISTICS
Recovery

Five parts of blank T cell culture supernatant were added with different concentrations of Anti-CD3 Antibody, and the T cell culture supernatant without Anti-CD3 Antibody was used as background to calculate the recovery rate. The range of the recovery rate is 81.3-102.5%, and the average recovery is 89.0%.

CD3 RECOVERY
  • Clinical and Translational Updates

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