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Your Position: Home > Kits > Pyrophosphatase > RES-A005

resDetect™ Pyrophosphatase ELISA Kit

For research use only.

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Product Details
Assay TypeSandwich-ELISA
AnalytePyrophosphatase
Format96T
ReactivityHuman
Regulatory StatusRUO
Sensitivity< 9.38 pg/mL
Standard Curve Range9.375 pg/mL-300 pg/mL
Assay Time3 hr 20 min
Suitable Sample TypeFor the quantitative determination of Pyrophosphatase in Cell Culture Supernatants.
Sample volume100 μL
Materials Provided
IDComponentsSize
RES005-C01Pre-coated Anti-Pyrophosphatase Antibody Microplate1 plate
RES005-C02Pyrophosphatase Standard100 μL
RES005-C03Biotin-Anti-Pyrophosphatase Antibody15 μg
RES005-C04Streptavidin-HRP50 μL
RES005-C0520xWashing Buffer 50 mL
RES005-C062xDilution Buffer50 mL
RES005-C07Substrate Solution12 mL
RES005-C08Stop Solution7 mL
  • Background
    Pyrophosphatase ELISA Kit(Residue Testing) is based on ELISA sandwich method and designed to detect and quantitative determine of Pyrophosphatase in mRNA preparation processing. It contains Pyrophosphatase and a pair of antibodies against the recombinant enzyme, 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 Pyrophosphatase concentrations. The specificity has been verified.
  • Application

    The kit is developed for the detection of Pyrophosphatase in mRNA drug products or semi-manufactures.

    It is for research use only.

  • Reconstitution
    Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • Storage
    The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.

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

    c) Add the Biotin-Anti-Pyrophosphatase 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 enzyme.

Typical Data Please refer to DS document for the assay protocol.
 Pyrophosphatase 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
Intra-Assay Statistics

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision, Intra-Assay Precision CV<15%.

 Pyrophosphatase 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<15%.

 Pyrophosphatase INTER-ASSAY STATISTICS
Recovery

Three Pyrophosphatase with different concentrations were tested to calculate the recovery rate.

 Pyrophosphatase RECOVERY
  • Background: Pyrophosphatase
    Pyrophosphatase catalyzes the hydrolysis of inorganic pyrophosphate to form orthophosphate, it can hydrolyze inorganic pyrophosphate generated with the reaction to avoid its inhibition of the reaction system. Pyrophosphatase is usually used to increase RNA yield in reverse transcription reactions. As a key raw material for in vitro transcription (IVT) of RNA, Pyrophosphatase needs to detect the residues as a protein component. This ELISA kit can be used to detect the residue of Pyrophosphatase in mRNA stock solution.
  • Clinical and Translational Updates

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