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Aav quantitation kit

Manufactured by Cell Biolabs
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The AAV quantitation kit is a tool for quantifying adeno-associated virus (AAV) particles. It provides a standardized method for determining the titer of AAV samples.

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Lab products found in correlation

Vivaspin 20 concentrator, by Sartorius (2 mentions) Aav dj 8 helper free expression system, by Cell Biolabs (1 mentions) Paav dj 8 vector, by Cell Biolabs (1 mentions) Paav mcs expression vector, by Cell Biolabs (1 mentions) 293aav cells, by Cell Biolabs (1 mentions)

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QuickTiter AAV Quantitation Kit (25 citations)

2 protocols using aav quantitation kit

1

Construction and Characterization of pAAV-hTCF21 Promoter Vector

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The pAAV-hTCF21 promoter vector was generated by cloning the 1013 bp human TCF21 (hTCF21) promoter fragment (Switchgear Genomics, Cat# S712020) into the ClaI and EcoR1 sites of the pAAV-MCS-Promoterless vector (Cell Biolabs, Cat# VPK-411). The FLAG-YAP(S127A) cDNA (Addgene 27370) was cloned to the EcoRI and BamHI sites of the pAAV-hTCF21 promoter vector. The GFP cDNA was cloned into the SalI and Hind III sites of the pAAV-hTCF21 promoter vector. Recombinant AAV was produced using the AAV-DJ/8 helper-free expression system (Cell Biolabs, Inc.) as described16 (link). Briefly, the recombinant pAAV-hTCF21 vector was co-transfected into 293AAV cell line along with pAAV-DJ/8 and pHelper in a 1:1:1 ratio using polyethylenimine. After 72 h of transfection, cells were harvested and purified by the iodixanol gradient/ultra-centrifugation method. The AAV fraction obtained from the gradient centrifugation was concentrated by the Vivaspin 20 concentrator (100 kDa cut-off, Sartorius, Germany). The virus titer was determined using the Cell Biolabs AAV quantitation kit (Cat. # VPK-145).
Francisco J., Zhang Y., Nakada Y., Jeong J.I., Huang C.Y., Ivessa A., Oka S., Babu G.J, & Del Re D.P. (2021). AAV-mediated YAP expression in cardiac fibroblasts promotes inflammation and increases fibrosis. Scientific Reports, 11, 10553.
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2

Recombinant AAV Vector Production

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The recombinant adeno-associated virus (AAV) vectors used to generate AAV-DJ/8-PPARα-WT or S280D were constructed by cloning the cDNA of PPARα-WT or S280D (mouse) from the pDC316-YFP-PPARα-WT or S280D vector into the pAAV-MCS expression vector (Cell Biolabs, Inc, #VPK-410) downstream of the CMV promoter with BamHI and SalI. 293AAVcells (Cell Biolabs, Inc.) were co-transfected with the recombinant AAV vectors, pAAV-DJ/8 vector, and helper plasmid in a 1:1:1 ratio using polyethylenimine (PEI) at the AAV core, Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers University, Newark, NJ (Grimm et al., 2008 (link)). The recombinant AAV produced was purified by the iodixanol gradient/ultra-centrifugation method, and the AAV fraction was concentrated using a VIVASPIN 20 concentrator (100 kDa cut-off, Sartorius, Germany). The virus titer was determined using the Cell Biolabs AAV quantitation kit (Cat. # VPK-145). To administer recombinant AAVs, doses of 2×1011 vector genome per mouse were injected intravenously via the jugular vein of C57BL/6J wild-type mice.
Nakamura M., Liu T., Husain S., Zhai P., Warren J.S., Hsu C.P., Matsuda T., Phiel C.J., Cox J.E., Tian B., Li H, & Sadoshima J. (2019). Glycogen Synthase Kinase-3α Promotes Fatty Acid Uptake and Lipotoxic Cardiomyopathy. Cell metabolism, 29(5), 1119-1134.e12.
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