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resDetect™ Human Interleukin-2 (IL-2) ELISA Kit (Residue Testing)

For research use only.

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Product Details
Assay TypeSandwich-ELISA
AnalyteIL-2
Format96T
ReactivityHuman
Regulatory StatusRUO
Sensitivity< 39.06 pg/mL
Standard Curve Range39.06 pg/mL-2500 pg/mL
Assay Time2 hr 50 min
Suitable Sample TypeFor the quantitative determination of human IL-2 in Cell Culture Supernatants, Plasma, Serum.
Sample volume100 uL
Materials Provided
IDComponentsSize
CRS003-C01Pre-coated Anti-IL-2 Antibody Microplate1 plate
CRS003-C02Human IL-2 Standard20 μg
CRS003-C03Biotin-Anti-IL-2 Antibody50 μL
CRS003-C04Streptavidin-HRP50 μL
CRS003-C0510xWashing Buffer 50 mL
CRS003-C062xDilution Buffer50 mL
CRS003-C07Substrate Solution12 mL
CRS003-C08Stop Solution7 mL
  • Background
    Human Interleukin-2 (IL-2) ELISA Kit (Residue Testing) is based on ELISA sandwich method and designed to detect and quantitative determine of GMP human IL-2 in samples from CAR-T product preparation processing. It contains GMP human IL-2 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 IL-2 (ACROBiosystems, cat#GMP-L02H14) concentrations. The specificity has been verified.
  • Application

    Human Interleukin-2 (IL-2) ELISA Kit (Residue Testing) was developed for the detection and quantitative determination of GMP human IL-2 in samples from CAR-T product preparation processing where IL-2 is used as an immunotherapy agent.

    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 IL-2. The kit consists of Pre-coated Anti-IL-2 Antibody Microplate and Human IL-2 Standard and Biotin-Anti-IL-2 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 IL-2 as standard. The samples and standard are diluted by Dilution Buffer.

    c) Wash the plate and add the Biotin-Anti-IL-2 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.
 IL-2 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 IL-2 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

 IL-2 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%.

 IL-2 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%.

 IL-2 INTER-ASSAY STATISTICS
Recovery

Three parts of blank serum were added with different concentrations of human IL-2, and the serum without human IL-2 was used as background to calculate the recovery rate. The range of the recovery rate is 83.9-98.2%, and the average recovery is 90.2%.

 IL-2 RECOVERY
  • Background: IL-2
    IL2,TCGF,lymphokine,Interleukin 2
    Interleukin-2 (IL-2) is an interleukin, a type of cytokine immune system signaling molecule, which is a leukocytotrophic hormone that is instrumental in the body's natural response to microbial infection and in discriminating between foreign (non-self) and self. IL-2 mediates its effects by binding to IL-2 receptors, which are expressed by lymphocytes, the cells that are responsible for immunity. Mature human IL-2 shares 56% and 66% aa sequence identity with mouse and rat IL-2, respectively. Human and mouse IL-2 exhibit crossspecies activity. The receptor for IL-2 consists of three subunits that are present on the cell surface in varying preformed complexes. IL-2 is also necessary during T cell development in the thymus for the maturation of a unique subset of T cells that are termed regulatory T cells (T-regs). After exiting from the thymus, T-Regs function to prevent other T cells from recognizing and reacting against "self antigens", which could result in "autoimmunity". T-Regs do so by preventing the responding cells from producing IL-2. Thus, IL-2 is required to discriminate between self and non-self, another one of the unique characteristics of the immune system.
  • Clinical and Translational Updates

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