AAVpro Purification Kit (All Serotypes)
AAVpro Purification Kit (All Serotypes)
Brand: Takara Bio.
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AAVpro Purification Kit (All Serotypes)
AAVpro Purification Kit (All Serotypes)
The AAVpro Purification Kit (All Serotypes) is a four-prep kit that can be used for purifying AAV of any serotype without the need for ultracentrifugation. The purification yields highly pure AAV particles that can be used directly for in vivo transduction or transduction of cultured cells. The kit includes Cryonase Cold-Active Nuclease.
Overview
- Kits include all reagents needed to purify AAV particles from vector-producing cells
- Purification yields highly pure AAV particles that can be used directly for in vivo transduction or transduction of cultured cells
- AAVpro Purification Kit (All Serotypes) is a four-prep kit which includes Cryonase Cold-Active Nuclease
Summary of the AAV particle purification protocol using the AAVpro Purification Kit (All Serotypes)
Summary of the AAV particle purification protocol using the AAVpro Purification Kit (All Serotypes).
Purity of viral particles collected using the AAVpro Purification Kit (All Serotypes)
Purity of viral particles collected using the AAVpro Purification Kit (All Serotypes). AAV viral particles of each serotype carrying the fluorescent protein ZsGreen1 were prepared from producer cells cultured in five T225 flasks. The AAV solution was purified using this product, and viral titer was measured using the AAVpro Titration Kit (for Real Time PCR) Ver. 2. SDS-PAGE was performed on each sample, with 1 x 109 vector genomes loaded per lane. For all serotypes, the AAV capsid proteins (VP1, VP2, and VP3) were observed to be the major bands present, confirming the purity of the AAV viral particles.
Infection capacity of AAV particles purified with the AAVpro Purification Kit (All Serotypes)
Infection capacity of AAV particles purified with the AAVpro Purification Kit (All Serotypes). Infectivities of AAV particles of various serotypes were evaluated. Purified AAV particles carrying the fluorescent protein ZsGreen1 were used for infection at 5,000 vector genomes per cell (serotypes 1, 2, 3, and 6) or 50,000 vector genomes per cell (serotype 5). Flow cytometry was performed three days later. Results indicate the AAV particles purified using this kit were able to infect the cell lines tested.
Gene transduction of iPSC-derived cardiomyocytes using AAV1, 2, 5, and 6.
Gene transduction of iPSC-derived cardiomyocytes using AAV1, 2, 5, and 6. AAV1, 2, 5, and 6 vectors carrying the ZsGreen1 gene were purified with the AAVpro Purification Kit Midi (All Serotypes) kit. Each purified vector was used for viral transduction of iPSC-derived cardiomyocytes at 500,000 vector genomes per cell. After 72 hr of infection, ZsGreen1 fluorescence was analyzed by microscopy, with an exposure time of 0.2 sec. AAV5 was additionally exposed at 0.5 sec. NC = negative control.
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Summary of the AAV particle purification protocol using the AAVpro Purification Kit (All Serotypes)
Summary of the AAV particle purification protocol using the AAVpro Purification Kit (All Serotypes).
Purity of viral particles collected using the AAVpro Purification Kit (All Serotypes)
Purity of viral particles collected using the AAVpro Purification Kit (All Serotypes). AAV viral particles of each serotype carrying the fluorescent protein ZsGreen1 were prepared from producer cells cultured in five T225 flasks. The AAV solution was purified using this product, and viral titer was measured using the AAVpro Titration Kit (for Real Time PCR) Ver. 2. SDS-PAGE was performed on each sample, with 1 x 109 vector genomes loaded per lane. For all serotypes, the AAV capsid proteins (VP1, VP2, and VP3) were observed to be the major bands present, confirming the purity of the AAV viral particles.
Infection capacity of AAV particles purified with the AAVpro Purification Kit (All Serotypes)
Infection capacity of AAV particles purified with the AAVpro Purification Kit (All Serotypes). Infectivities of AAV particles of various serotypes were evaluated. Purified AAV particles carrying the fluorescent protein ZsGreen1 were used for infection at 5,000 vector genomes per cell (serotypes 1, 2, 3, and 6) or 50,000 vector genomes per cell (serotype 5). Flow cytometry was performed three days later. Results indicate the AAV particles purified using this kit were able to infect the cell lines tested.
Gene transduction of iPSC-derived cardiomyocytes using AAV1, 2, 5, and 6.
Gene transduction of iPSC-derived cardiomyocytes using AAV1, 2, 5, and 6. AAV1, 2, 5, and 6 vectors carrying the ZsGreen1 gene were purified with the AAVpro Purification Kit Midi (All Serotypes) kit. Each purified vector was used for viral transduction of iPSC-derived cardiomyocytes at 500,000 vector genomes per cell. After 72 hr of infection, ZsGreen1 fluorescence was analyzed by microscopy, with an exposure time of 0.2 sec. AAV5 was additionally exposed at 0.5 sec. NC = negative control.
