AAV2 VP1 + VP2 + VP3, recombinant proteins, set
Analysis of AAV2 VP1 + VP2 + VP3, recombinant proteins (Cat. No. 72001) with various stoichiometric ratios by Simple WesternTM, a CE-immunoassay platform from ProteinSimple, a Bio-Techne brand. Capsid proteins were detected using an anti-AAV VP1/VP2/VP3 mouse monoclonal antibody (Cat. No. 690058).
Western blot analysis was performed on different molar ratios of VP1:VP2:VP3 (1:1:10, 2:2:5, 1:1:5, 3:3:10 and 3:3:3) compared to 1E+10 denatured AAV2 capsids. The capsids were denatured for 10 min at 95°C. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-AAV VP1/VP2/VP3 mouse monoclonal, B1 (Cat. No. 690058) was diluted in blocking buffer (antibody concentration 0.5 µg/ml) and incubated for 1 h at RT. The secondary antibody goat anti-mouse IgG HRP was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Please note that various amounts of total protein were loaded in each lane.
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).
Undiluted = 100 µg/ml, pre-diluted = 10 µg/ml
Analysis of AAV2 VP1 + VP2 + VP3, recombinant proteins (Cat. No. 72001) with various stoichiometric ratios by Simple WesternTM, a CE-immunoassay platform from ProteinSimple, a Bio-Techne brand. Capsid proteins were detected using an anti-AAV VP1/VP2/VP3 mouse monoclonal antibody (Cat. No. 690058).
Western blot analysis was performed on different molar ratios of VP1:VP2:VP3 (1:1:10, 2:2:5, 1:1:5, 3:3:10 and 3:3:3) compared to 1E+10 denatured AAV2 capsids. The capsids were denatured for 10 min at 95°C. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-AAV VP1/VP2/VP3 mouse monoclonal, B1 (Cat. No. 690058) was diluted in blocking buffer (antibody concentration 0.5 µg/ml) and incubated for 1 h at RT. The secondary antibody goat anti-mouse IgG HRP was also diluted in blocking buffer (antibody concentration 0.2 µg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Please note that various amounts of total protein were loaded in each lane.
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).
Undiluted = 100 µg/ml, pre-diluted = 10 µg/ml
• Useful for ratio analysis
• Precise molar concentration
• Suitable for WB, dot blot, CE, SDS PAGE
Included products:
Product description
| Quantity | 10 µg each protein |
|---|---|
| Application | Capillary electrophoresis (CE), Dot blot, SDS PAGE, WB |
| Purification | Ni-NTA chromatography |
| Storage | -80°C |
| Intended use | Research use only |
| Concentration | 100 µg/ml (VP1: 1.19 µM, VP2: 1.45 µM, VP3: 1.61 µM) |
| Formulation | Liquid, 6 M urea in PBS |
| Source | Escherichia coli |
| Molecular weight | VP1: 84.1 kDa, VP2: 68.9 kDa, VP3: 62.2 kDa (calculated Mw from aa sequence) |
| Purity | > 90% (determined by SDS PAGE) |
| Product description | N-terminal His-tagged (MGSSHHHHHHSSGLVPRGSH) recombinant AAV2 capsid proteins VP1 + VP2 + VP3 |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Capillary electrophoresis (CE) | Assay dependent |
| 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.
This set of recombinant AAV2 VP1, VP2 and VP3 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 AAV2 capsid proteins are available as set (Cat. No. 72001) or as individual proteins (Cat. No. 640823, 640824, 640825).
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