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AAV8 VP3, recombinant protein

SDS PAGE analysis to evaluate the purity of the AAV8 VP3 (Cat. No. 640841). To perform SDS PAGE analysis, 1 µg of protein was diluted in 10 µl PBS and sample buffer and denatured at 95°C for 5 min. The sample was loaded onto a 4-20% gradient gel (40 min at 200 V). Afterwards, the gel was stained for 1 h at RT with Coomassie solution and destained with water. The purity of AAV8 VP3 is > 95%.
Western blot analysis of recombinant AAV8 VP3 (Cat. No. 640841) with B1 antibody (Cat. No. 690058) and ECL detection. Western blot analysis was performed on different amounts of recombinant AAV-VP3 ranging from 7 ng to 34 ng. The PVDF membrane was blocked with 5% milk in PBST for 1 h at RT. The primary antibody anti-AAV VP1/VP2/VP3, B1 (Cat. No. 690058) was diluted in blocking buffer (antibody concentration 500 ng/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG HRP was also diluted in blocking buffer (antibody concentration 200 ng/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using Pierce™ ECL Western Blotting Substrate.
SDS PAGE analysis to evaluate the purity of the AAV8 VP1, VP2 and VP3 (Cat. No. 72008). To perform SDS PAGE analysis, 1 µg of protein was diluted in 10 µl PBS and sample buffer and denatured at 95°C for 5 min. The samples were loaded onto a 4-20% gradient gel (40 min at 200 V). Afterwards, the gel was stained for 1 h at RT with Coomassie solution and destained with water. The purity of AAV8 VP1, VP2 and VP3 is > 95%.

Western blot analysis of recombinant AAV2 VP proteins (Cat. No. 72001), recombinant AAV5 VP proteins (Cat. No. 72005), recombinant AAV6 VP proteins (Cat. No. 72006), recombinant AAV8 VP proteins (Cat. No. 72008) and recombiant AAV9 VP proteins (Cat. No. 72009) with B1 antibody (Cat. No. 690058). Western blot analysis was performed on the precise molar ratio of 1:1:10 (13.5 ng VP1:11 ng VP2:100 ng VP3) combined in one lane. The PVDF membrane was blocked with 5% milk in PBST for 1 h at RT.
The primary antibody anti-AAV VP1/VP2/VP3, B1 (Cat. No. 690058) was diluted in blocking buffer (antibody concentration 500 ng/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG HRP was also diluted in blocking buffer (antibody concentration 200 ng/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using Pierce™ ECL Western Blotting Substrate.


Pipetting scheme for western blot analysis using a mix of the AAV8 capsid proteins (Cat. No. 640839, 640840, 640841). To create a VP mixture with the molar ratio 1:1:10 (VP1:VP2:VP3), please pre-dilute VP1 and VP2 1:10 to yield a final concentration of 10 µg/ml (green table). Pipette the pre-diluted VP1 and VP2 proteins and mix them with the undiluted VP3 protein in your sample buffer and water (blue table). The example with 2x and 3x sample buffer and the required volumes are indicated in the pipetting scheme. Thus, in one lane, 10 µl of the VP mix can be loaded onto the SDS PAGE and analyzed by Western blot using the B1 antibody (Cat. No. 690058, Cat. No. 61058-488, Cat. No. 61058-647).
Undiluited = 100 µg/ml, pre-diluted = 10 µg/ml
Western blot analysis of recombinant AAV8 capsid proteins (Cat. No. 640839, 640840, 640841)  and denatured AAV8 capsids with B1 antibody (Cat. No. 690058). Western blot analysis was performed on the precise molar ratio of 1:1:10 (VP1:VP2:VP3) either in separate lanes or combined in one lane and on 3.3E+09 denatured AAV8 capsids. The PVDF membrane was blocked with 5% milk in PBST for 1 h at RT. The primary antibody anti-AAV VP1/VP2/VP3, B1 (Cat. No. 690058) was diluted in blocking buffer (antibody concentration 500 ng/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG HRP was also diluted in blocking buffer (antibody concentration 200 ng/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using Pierce™ ECL Western Blotting Substrate.


Pipetting scheme for western blot analysis using the AAV8 capsid proteins (Cat. No. 640839, 640840, 640841) in separate lanes. To analyze the molar ratio of 1:1:10, it is recommended to load VP1, VP2 and VP3 as described in the pipetting scheme above. Therefore, the indicated volumes of the proteins (concentration 100 µg/ml) should be diluted with the appropriate amount of sample buffer and distilled water. 10 µl of each solution can be separately loaded onto the SDS PAGE and analyzed by Western blot using the B1 antibody (Cat. No. 690058, Cat. No. 61058-488, Cat. No. 61058-647).
Cat. No: 640841

Prices excl. tax

Available, delivery time: 1-10 days

$342.00*

Datasheet
Key Features
•    Recombinant AAV8 capsid protein VP3
•    Useful in combination with AAV8 VP1 and VP2 for ratio analysis (also available as set, Cat. No. 72008)
•    Precise molar concentration
•    Suitable for WB, dot blot, SDS PAGE

Not available in China and 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
Quantity 10 µg
Application Dot blot, SDS PAGE, WB
Purification Ni-NTA chromatography
Storage -80°C
Intended use Research use only
Concentration 100 µg/ml (1.61 µM)
Formulation Liquid, 6 M urea in PBS
Source Escherichia coli
Molecular weight 62.1 kDa (calculated Mw from aa sequence)
Purity > 95% (determined by SDS PAGE)
Product description

N-terminal His-tagged (MGSSHHHHHHSSGLVPRGSH) recombinant AAV8 capsid protein VP3
Applications
Tested applications Tested dilutions
Dot Blot 100 ng, depending on primary antibody and detection method
SDS PAGE 1 µg
Western Blot (WB) 5-20 ng, depending on primary antibody and detection method
Background

The AAV capsid consists of three capsid proteins, i.e. VP1, VP2 and VP3, which differ in their N-terminus and encapsulate the genomic ssDNA. In native virus particles, the three proteins form subunits with a ratio of 1:1:10 (VP1:VP2:VP3), in a total number of 60 subunits per capsid.
The recombinant AAV8 VP3 protein in combination with recombinant AAV8 VP1 (Cat. No. 640839) and recombinant AAV8 VP2 (Cat. No. 640840) can be used to create a mixture with the precise molar ratio of 1:1:10 to compare the protein composition of the viral capsid in your sample by protein detection methods, e.g. western blot.
All three recombinant AAV8 capsid proteins are available as set (Cat. No. 72008) or as individual proteins (Cat. No. 640839, 640840, 640841).

Note: please find an example how to prepare western blot samples in the pipetting scheme below. Aliquots of the remaining samples can be stored at -80°C for reuse.
Downloads
File
Category
Size
Filetype
File Poster AAV VP protein standard
Category Poster
Size 1.44 MB
Filetype pdf
Datasheet
MSDS
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