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Your Position: Home > Kits > AAV9 > AAV-A009H

AAV9 Titration ELISA Kit (Fast)

For research use only.

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    Materials Provided
    IDComponentsSize
    AAV09H-C01Pre-coated Anti-AAV9 Antibody Microplate1 plate
    AAV09H-C02AAV9 Standard3.80E+11 capsids
    AAV09H-C03HRP-Anti-AAV9 Antibody20 μg
    AAV09H-C0410×Washing Buffer 50 mL
    AAV09H-C052×Dilution Buffer50 mL
    AAV09H-C06Substrate Solution12 mL
    AAV09H-C07Stop Solution7 mL
  • Background
    Adeno-associated virus (AAV) has become one of the most important gene vectors in the field of gene therapy due to its long-term expression, low toxicity, low immunogenicity, and high tissue specificity.Most successful AAV gene therapies for preclinical and clinical studies are limited to natural serotypes, but the presence of neutralizing antibodies against AAV remains a significant barrier to systemic delivery.
  • Application

    AAV9 Titration ELISA Kit was developed for the detection and quantitative determination of AAV9 capsid titration in AAV gene therapy product preparation processing.

    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 sandwich-ELISA format, providing a rapid detection of AAV9 capsid. The kit consists of Pre-coated Anti-AAV9 Antibody Microplate and AAV9 Standard,HRP-Anti-AAV9 Antibody and buffers.

    Your experiment will include 5 simple steps:

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

    b) Add your sample to the plate, take the AAV9 capsid as Control sample. The samples and Control sample are diluted by Dilution Buffer.

    c) Add the HRP-Anti-AAV9 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 prior to statistical analysis. The OD Value reflects the amount of protein bound.

Typical Data Please refer to DS document for the assay protocol.
 AAV9 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 AAV9 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

 AAV9 DILUTION LINEARITY
Intra-Assay Statistics

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

 AAV9 INTRA-ASSAY STATISTICS
Inter-Assay Statistics

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

 AAV9 INTER-ASSAY STATISTICS
Recovery

Three AAV9 with different concentrations were tested to calculate the recovery rate.

 AAV9 RECOVERY
Specificity

The specific binding of AAV-A009H to AAV1/2/3/5/6/8/9/dj serotypes was detected. The results showed that AAV-A009H only binds specifically to AAV9.

 AAV9 SPECIFICITY

The specific binding of AAV-A009H to native/denatured AAV9 was detected. The results showed that AAV-A009H only binds specifically to native AAV9. (Denatured AAV9: 95℃ for 5min)

 AAV9 SPECIFICITY
  • Background: AAV9
    The adeno-associated virus (AAV) is a small (25 nm), non-enveloped virus of the parvoviridae family, including 12 different AAV serotypes. In the parvoviridae family it belongs to the genus dependoparvovirus, because it needs the presence of a helper virus for replication and assembly. The icosahedral AAV capsid composed of the capsid proteins VP1, VP2 and VP3 contains a linear, single-stranded DNA genome of 4.7 kb.
  • Clinical and Translational Updates

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