Anti-AAV VP1/VP2/VP3

Dot blot with anti-AAV VP1/VP2/VP3 AFDyeTM 647 antibody (Cat. No. 61058-647, 0.1 µg/ml) using native and denatured capsids of the serotypes AAV6 and AAV8.

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Binding Sites and Reactivity

The monoclonal antibody B1, as well as the polyclonal serum VP51 react with the three AAV capsid proteins VP1, VP2 and VP3. Both antibodies were raised against the AAV2 capsid proteins and are suitable for application in western blot analysis due to their reactivity with linear epitopes present in the dentaured protein sequence of all three proteins. As you can see on the dotplot, PROGEN's B1-antibodie does not detect assembled capsids. 

Epitope mapping experiments (Wobus et al., 2000) identified aa726 to aa733 (C-terminus; common to all 3 VP proteins) as the specific binding region for the monoclonal B1 antibody. The antibodies are useful for the characterization of different stages of AAV infection, the determination of the presence and identity of VP1/VP2/VP3 as well as the analysis of their expression levels and ratios during vector production. For more information regarding binding sites please read the publication below.

WB with anti-AAV VP1/VP2/VP3 antibody (Cat. No. 61058, 1:250) using AAV2 capsids as sample

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Wobus, C. E. et al. Monoclonal antibodies against the adeno-associated virus type 2 (AAV-2) capsid: epitope mapping and identification of capsid domains involved in AAV-2-cell interaction and neutralization of AAV-2 infection. J. Virol. 74, 9281–93 (2000).

The AFDye™

The use of a AFDyeTM labeled antibody enables the detection of proteins without the need of a secondary antibody which saves valuable time during processing. Theses Dyes are structurally similar to Alexa Fluor™ which is a trademark of Thermo Fisher Scientific. PROGEN offers two different AFDyes™ conjugates: AFDye™647 and AFDye™488.

AFDye™647

The AFDye™ 647 is a bright, far-red-fluorescent dye ideally suited for the 633 nm or 647 nm laser lines (excitation/emission 648/671 nm). It is structurally similar to Alexa Fluor® 647, and can be used with a common Cy5 filter set. A significant advantage to using long wavelength dyes such as Cy5 or AFDye™ 647 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum.

AFDye™488

The AFDye™ 488 is a green emitting dye, structurally and spectrally equivalent to Alexa Fluor® 488 and therefore ideally suited for the 488 nm laser line (excitation/emission 494/517 nm). It can be used with a common FITC filter set.