AAV5 standard material (eGFP)

Cat. No: 66V051

Available, delivery time 1-7 days

$647.00*

Key Features
  • Fully assembled AAV5 capsids filled with an eGFP reporter gene controlled by a CMV promoter
  • Serotype-specific conformational epitopes
  • Reliable positive control for dot blot, western blot, ELISA, PCR and cell-based assays
  • Quality controlled for purity, filling grade, aggregation, total capsid and genome titer
  • Alignment to internal reference standard material
Not available in the Russian Federation.
For US shipment, the packages are only sent out for delivery on Mondays and Tuesdays, in the EU from Monday until Wednesday.
Product description
Filling grade > 70% full capsids, please find the lot-specific filling grade on the CoA
Quantity 100 µl
Application Calibration of instruments e.g. mass photometry, Cell-based assay, Dot blot, ELISA, WB, ddPCR, qPCR
Purification Affinity Chromatography (POROS CaptureSelect AAVX Affinity Resin, Thermo Fisher Scientific), Iodixanol gradient centrifugation for removal of empty capsids using OptiPrep (Cat. No. 1893, PROGEN)
Storage Up to 2 weeks: 2-8°C; long term storage in aliquots at -80°C; avoid > 5 freeze/thaw cycles
Intended use Research use only
Concentration

> 1.0E+12 vg/ml; please find the lot-specific concentration (total capsid and viral genome titer) on the CoA

Formulation PBS + 0.014% Tween20 + 1 mM MgCl2 + 2.5 mM KCl
Source Produced in HEK293T cells
Purity > 95% by SDS-PAGE and silver stain
Quality check

Total capsid titer was assigned using AAV5 ELISA kits (PRAAV5 and PRAAV5XP, calibrated with internal reference material) and viral genome titer by qPCR using eGFP primers; QC included analysis of filling grade, purity, aggregation, and endotoxin testing.

Transfer Vector pAAV-CMV-eGFP-WPRE
Packaging Plasmid pDP5rs
Endotoxin < 1.0 EU/ml (detection limit 1.0 EU/ml)
Note Please centrifuge before opening to ensure complete recovery of vial contents; aliquoting and repeated freeze/thaw cycles can lead to a drop in titer
Applications
Tested applications Tested dilutions
Dot Blot Depending on primary antibody and detection method
ELISA As a positive control in ELISA, dilute in ASSB 1x (provided with PROGEN’s AAV5 ELISA), analysis at least in duplicates is recommended
Cell-based Assay Depending on the experimental setup
PCR As standard or positive control in qPCR or ddPCR - concentration depending on experimental setup
Western Blot (WB) Depending on primary antibody and detection method
Background

Our AAV5 standard material consists of fully assembled AAV5 capsids with an eGFP reporter gene controlled by a CMV promotor. Final concentrations (total capsids and viral genome titer) are lot-specific and can be found on the CoA. 

The AAV5 standard material is provided with a titer above 1.0E+12 viral genomes/ml in a liquid formulation. Since the buffer does not contain any stabilizing proteins or dyes, the capsids can be used in various applications, including dot blot, western blot, ELISA, qPCR, ddPCR and cell-based assays. Our material contains only a very low remaining amount of iodixanol allowing DLS-SLS-UV/Vis analysis. The lot-specific titers were assigned using AAV5 ELISA (PRAAV5 and PRAAV5XP) according to our internally established standard material* and PCR using eGFP primers. Our comprehensive quality control ensures well-characterized capsid material, which can be implemented as reference material in a variety of assays to prove the validity of the corresponding assay.

PROGEN provides eGFP-filled AAV standard material for the AAV serotypes 1, 2, 3, 5, 6, 8, 9, rh10, and rh74.

*Our internal standard material for each serotype was characterized according to the protocol described in our poster Developing Reliable AAV Standards for ELISA (available in the downloads tab). Data on the establishment of standard material for specific serotypes can be found as part of the performance data for the corresponding ELISAs or can be provided upon request.

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FAQs

Please have a look for the table below to see the combination of plasmids used for the production of our AAV standard material: 

Serotype Transfer vector Packaging plasmid Helper plasmid
AAV1 pAAV-CMV-eGFP-WPRE  pDP1rs 
AAV2 pAAV-CMV-eGFP-WPRE pDP2rs 
AAV3 pAAV-CMV-eGFP-WPRE  pRep2-Cap3  pHelper 
AAV5 pAAV-CMV-eGFP-WPRE  pDP5rs 
AAV6 pAAV-CMV-eGFP-WPRE  pRep2-Cap6  pHelper 
AAV8 pAAV-CMV-eGFP-WPRE  pRep2-Cap8  pHelper 
AAV9 pAAV-CMV-eGFP-WPRE  pRep2-Cap9  pHelper 
AAVrh10 pAAV-CMV-eGFP-WPRE  pRep2-Caprh10  pHelper 
AAVrh74 pAAV-CMV-eGFP-WPRE  pRep2-Caprh74  pHelper 

High virus titers > 1.0E+12 vg/ml and liquid formulation (PBS with 0.014% Tween, 1 mM MgCl2 and 2.5 mM KCl) allow the use as a reference material in a variety of applications e.g. PCR, BLI, SEC-MALS, mass photometry, infectivity and many more.

The AAV capsids titer is measured by our PROGEN ELISAs and the genomic titer is measured by qPCR using eGFP primers.

Our AAV standard material contains the most commonly used and well known combination of an eGFP transgene with a CMV promoter

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