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resDetect™ Human Interleukin-6 (IL-6) ELISA Kit (Residue Testing)

For research use only.

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Product Details
Assay TypeSandwich-ELISA
AnalyteIL-6
Format96T
ReactivityHuman
Regulatory StatusRUO
Sensitivity< 2.34 pg/mL
Standard Curve Range2.34 pg/mL-150 pg/mL
Assay Time2 hr 50 min
Suitable Sample TypeFor the quantitative determination of human IL-6 in Cell Culture Supernatants, Plasma, Serum.
Sample volume100 uL
Materials Provided
IDComponentsSize
CRS005-C01Pre-coated Anti-IL-6 Antibody Microplate1 plate
CRS005-C02Human IL-6 Standard20 μg
CRS005-C03Biotin-Anti-IL-6 Antibody50 μL
CRS005-C04Streptavidin-HRP50 μL
CRS005-C0510xWashing Buffer 50 mL
CRS005-C062xDilution Buffer50 mL
CRS005-C07Substrate Solution12 mL
CRS005-C08Stop Solution7 mL
  • Background
    Human Interleukin-6 (IL-6) ELISA Kit (Residue Testing) is based on ELISA sandwich method and designed to detect and quantitative determine of GMP human IL-6 in samples from CAR-T product preparation processing. It contains GMP human IL-6 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-6 (ACROBiosystems, cat#GMP-L06H27) concentrations. The specificity has been verified.
  • Application

    Human Interleukin-6 (IL-6) ELISA Kit (Residue Testing) was developed for the detection and quantitative determination of GMP human IL-6 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 IL-6. The kit consists of Pre-coated Anti-IL-6 Antibody Microplate and Human IL-6 Standard and Biotin-Anti-IL-6 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-6 as standard. The samples and standard are diluted by Dilution Buffer.

    c) Wash the plate and add the Biotin-Anti-IL-6 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-6 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-6 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

 IL-6 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-6 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-6 INTER-ASSAY STATISTICS
Recovery

Three parts of blank serum were added with different concentrations of human IL-6, and the serum without human IL-6 was used as background to calculate the recovery rate. The range of the recovery rate is 90.6-114.8%, and the average recovery is 101.2%.

 IL-6 RECOVERY
  • Background: IL-6
    IL6,Interleukin-6,BSF2,HSF,IFNB2
    Interleukin 6 (IL-6) is also known as HGF, BSF2,HSF, IFNB2 and IL-6, originally identified as a B cell differentiation factor, is a multifunctional cytokine that regulates immune responses, hematopoiesis, acute phase responses, and inflammatory reactions.It is secreted by T cells, macrophages , monocytes, fibroblasts,endothelial cells,et.al. to stimulate immune response to trauma, especially burns or other tissue damage leading to inflammation. Interleukin 6 has been shown to interact with interleukin-6 receptor and glycoprotein. IL-6 is relevant to many disease processes such as diabetes,atherosclerosis, depression,Alzheimer's Disease,systemic,lupus erythematosus,prostate cancer and rheumatoid arthritis. Advanced/metastatic cancer patients have higher levels of IL-6 in their blood.Hence there is an interest in developing anti-IL-6 agents as therapy against many of these diseases.
  • Clinical and Translational Updates

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