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AAV-9 Titration ELISA




 

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Product Code: PRAAV9


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Product Details Protocol
 
Product Code: PRAAV9
Product: AAV-9 Titration ELISA
Description: Enzyme-linked Immunosorbant Assay (ELISA) for the Quantitative Determination of AAV Serotype 9 Particles in Cell Culture Supernatants and Purified Virus Preparations.

Unique microtiter plate enzyme immunoassay for the quantitation of intact AAV9 wt virions, AAV9 recombinant virions or assembled and intact empty capsids of human Adeno-associated Virus 9. The capture-antibody detects a conformational epitope not present on unassembled capsid proteins.
Background: Adeno-associated viruses (AAV) are non- pathogenic ssDNA viruses, which are a subject of intense studies as viral vectors for gene therapy. The virus transduces a variety of dividing and non- dividing cells showing long-term gene expression with low cellular immune response. AAV has been used in several clinical trials (e.g. FIX, CFTR, Parkinson’s, Canavan disease) showing no serious vector-related adverse effects. Methods for the characterization of AAV preparations currently include titration ELISA, real-time PCR, DNA dot blot, determination of transducing units, infectious center assay, SDS-PAGE or electron microscopy. Immunotitration by ARP's AAV9 Titration ELISA offers a fast, sensitive and reproducible method for titration of intact AAV9 wt virions, AAV9 recombinant virions or assembled and intact empty AAV9 capsids.
Principle: The assay is based on the sandwich ELISA technique. A monoclonal antibody specific for a conformational epitope on assembled AAV9 capsids (ADK9) is coated onto strips of a microtiter plate and is used to capture AAV9 particles from the specimen. Captured AAV particles are detected in two steps. First, a biotin-conjugated monoclonal antibody to AAV9 (ADK9) is bound to the immune complex. Second, a streptavidin peroxidase conjugate reacts with the biotin molecules. Addition of substrate solution results in a color reaction, which is proportional to the amount of specifically bound viral particles.

The absorbance is measured photometrically at 450 nm. The provided Kit Control contains an AAV9 particle preparation of empty capsids. Two-fold serial dilutions of the material result in a typical titration curve. The curve allows the quantitative determination of samples of an unknown particle titer.
Storage: 2-8C
Intended Use: For Research Use Only
This product is sold for laboratory research use or further manufacturing only and should not be used for human therapeutic or diagnostic applications. The information presented is believed to be accurate; however, said information and products are offered without warranty or guarantee since the ultimate conditions of use and the variability of the materials treated are beyond our control. Nothing disclosed herein is to be construed as a recommendation to use our products in violation of any patents. Under no circumstances shall ARP American Research Products, Inc. be liable for damages, whether consequential, compensatory, incidental or special, strict liability or negligence, breach of warranty or any other theory arising out of the use of the products available from ARP American Research Products, Inc. Nothing contained herein warrants that the use of the products will not infringe on the claims of any patents covering the product itself or the use thereof in combination with other products or in the operation of any process.

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