Recombinant Mouse Anti-AAV VP1 Antibody scFv Fragment (A20)
CAT#: PSBL-390
Recombinant Mouse Antibody (A20) scFv Fragment is specific to AAV VP1, expressed in E. coli. This scFv fragment neutralizes AAV-2 and AAV-3B subsequent to primary receptor binding.
Published Data
IP
Figure 1 Influence of mutation of pore-forming amino acids on genome packaging.
(B) Capsids were immunoprecipitated (IP) from frozen-thawed lysates obtained from 293T cells transfected with a plasmid harboring an AAV2 wt or mutated genome and superinfected with Ad5 (MOI, 2) by protein A-Sepharose-bound antibody A20. Control (C) precipitations were also performed with a non-AAV-related monoclonal antibody. Samples were digested (+) or not digested (−) with DNase I to discriminate between capsid-associated and packaged DNAs. Coprecipitated genomes were isolated and electrophoresed on neutral agarose gels. Southern blotting was performed, and genomes were detected with a rep-specific probe. A 4.7-kb fragment was used as a marker (top panels, M). A portion of each immunoprecipitate was tested by Western blot analysis for the level of capsid recovery; an extract from Ad5- and AAV2-coinfected HeLa cells was used as a marker (bottom panels, M).
Bleker, S., Sonntag, F., & Kleinschmidt, J. A. (2005). Mutational analysis of narrow pores at the fivefold symmetry axes of adeno-associated virus type 2 capsids reveals a dual role in genome packaging and activation of phospholipase A2 activity. Journal of virology, 79(4), 2528-2540.
DB
Figure 2 Externalization of VP1/VP2 N termini during infection.
(B) Analysis of cell lysates. HeLa cells were incubated with wt AAV2 (10,000 particles/cell) for 30 min at 4°C and washed. Following incubation at 37°C, the cells were harvested and lysed by freezing-thawing at the indicated time points. Virus-containing supernatant was analyzed by a native immuno-dot blot using VP1-specific antibody A1, VP1/VP2, specific antibody A69, C-terminus-specific antibody B1, or capsid-specific antibody A20. Extracts of noninfected cells were used as a negative control (n). Signal intensities were measured with Image Quant TL software, and the ratios of A1/A20 and A69/A20 signal were determined to quantify exposure of N termini on capsids. Means ± standard deviations from three independent experiments are shown.
Sonntag, F., Bleker, S., Leuchs, B., Fischer, R., & Kleinschmidt, J. A. (2006). Adeno-associated virus type 2 capsids with externalized VP1/VP2 trafficking domains are generated prior to passage through the cytoplasm and are maintained until uncoating occurs in the nucleus. Journal of virology, 80(22), 11040-11054.
Neut
Figure 3 AAV2 enters the cytoplasm with exposed VP1/VP2 N termini.
(A) Extracellular neutralization of infection. AAV2 particles were preincubated with 100 μg/ml of VP1-specific antibody A1, VP1/VP2-specific antibody A69, C-terminus-specific antibody B1, or capsid-specific antibody A20. In a control experiment, no antibody (no Ab) was added. The samples were transferred to HeLa cells (2 × 10³ particles/cell), infected with Ad5 (MOI = 4), and incubated for 20 h. Early viral gene expression was analyzed by immunofluorescence with Rep-specific antibody 76.3. Cell nuclei were visualized with DAPI. Means ± standard deviations from two independent experiments are shown.
Sonntag, F., Bleker, S., Leuchs, B., Fischer, R., & Kleinschmidt, J. A. (2006). Adeno-associated virus type 2 capsids with externalized VP1/VP2 trafficking domains are generated prior to passage through the cytoplasm and are maintained until uncoating occurs in the nucleus. Journal of virology, 80(22), 11040-11054.
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Specifications
- Immunogen
- AAV Capsid protein VP1
- Host Species
- Mouse
- Derivation
- Mouse
- Type
- scFv
- Specificity
- Tested positive against native AAV VP1
- Species Reactivity
- AAV
- Clone
- A20
- Applications
- WB, ELISA, FuncS, IP, DB, Neut
Product Property
- Purity
- >95% by SDS-PAGE and HPLC analysis
- Storage
- Store the antibody (in aliquots) at -20°C. Avoid repeated freezing and thawing of samples.
Applications
- Application Notes
- The VP1 antibody has been reported in applications of Western Blot, Enzyme-linked Immunosorbent Assay, Functional Assay, Immunoprecipitation, Dot Blot, Neutralization.
For Neutralization: wt AAV2 particles (2.5 × 10⁸) were incubated with 100 μg/ml purified antibody A1, A69, B1, or A20 in a total volume of 5 μl (PBS) for 30 min at room temperature. Then, the samples were diluted in 1 ml medium and added in combination with Ad5 (MOI = 4) to 1.4 × 10⁵ HeLa cells grown overnight on coverslides. Rep expression was analyzed after 20 h as described above.
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Quick Delivery
The antibody was delivered promptly, right when we needed it for a critical experiment. The fast delivery saved us a lot of time and kept our project on track.
No Non-Specific Binding
Performed well without any non-specific binding, even when used under stringent conditions. It's great to have such a reliable tool for our viral detection assays.
Adaptable to Different Protocols
We've been able to adapt this antibody to various prototyping and experimental protocols without losing efficacy, which speaks volumes about its robustness and versatility.
Q&As
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How do I prevent antibody degradation during storage?
A: Store the antibody under conditions specified on the datasheet, typically at -20°C or lower. Avoid prolonged exposure to light and repeated freeze-thaw cycles.
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What are potential causes of weak signals in my assays?
A: Causes may include suboptimal antibody concentration, insufficient antigen availability, poor-quality secondary antibody, or inadequate detection system sensitivity.
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Can I use this antibody with automated staining systems?
A: Yes, many recombinant antibodies are compatible with automated staining systems, but initial validation on your specific system is recommended to ensure optimal performance.
View the frequently asked questions answered by Creative Biolabs Support.
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Downloadable Resources
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Product Notes
This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:
• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production
See more details about Hi-Affi™ recombinant antibody benefits.
Datasheet
MSDS
COA
Certificate of Analysis LookupTo download a Certificate of Analysis, please enter a lot number in the search box below. Note: Certificate of Analysis not available for kit components.
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|---|---|---|---|
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| CAT | Product Name | Application | Type |
|---|---|---|---|
| PFBL-131 | Recombinant Mouse Anti-HEV-C VP1 Antibody Fab Fragment (C3) | WB, Neut, FuncS | Fab |
| PFBZ-047 | Recombinant Mouse Anti-FMDV VP1 pentadecapeptide (YTTSTRGDLAHVTAT) Antibody Fab Fragment (4C4) | Neut, ELISA | IgG |
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| FAMAB-0138-CN-F(E) | Recombinant Human Anti-VP1 Antibody Fab Fragment (1418-1) | WB, ELISA, FC | Human Fab |
| HPAB-AP484-YC-F(E) | Human Anti-VP1 Recombinant Antibody; Fab Fragment (HPAB-AP484-YC-F(E)) | ELISA, FC, IF, IHC, Dot, WB, Neut | Human Fab |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| PFBW-136 | Recombinant Mouse Anti-AAV VP1 Antibody Fab Fragment (A20) | WB | Fab |
| PSBZ-047 | Recombinant Mouse Anti-FMDV VP1 pentadecapeptide (YTTSTRGDLAHVTAT) Antibody scFv Fragment (4C4) | Neut, ELISA | scFv |
| PSBW-074 | Recombinant Mouse Anti-HRV14 VP1 Antibody scFv Fragment (17-IA) | Neut | scFv |
| PSBC-075 | Recombinant Mouse Anti-HRV14 VP1 Antibody scFv Fragment (1-IA) | Neut | scFv |
| HPAB-AP484-YC-S(P) | Human Anti-VP1 Recombinant Antibody; scFv Fragment (HPAB-AP484-YC-S(P)) | ELISA, FC, IF, IHC, Dot, WB, Neut | Human scFv |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| MHC-CN1333 | PE-A*01:01/Virus VP1 (SADNNNSEY) MHC Tetramer | FCM, IHC | |
| MHC-CN1334 | APC-A*01:01/Virus VP1 (SADNNNSEY) MHC Tetramer | FCM, IHC | |
| MHC-CN1335 | FITC-A*01:01/Virus VP1 (SADNNNSEY) MHC Tetramer | FCM, IHC | |
| MHC-CN1336 | A*01:01/Virus VP1 (SADNNNSEY) MHC Tetramer | FCM, IHC | |
| MHC-CN1337 | PE-A*02:01/Virus VP1 (ILMWEAVTL) MHC Tetramer | FCM, IHC |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| HPAB-0504-YJ-VHH | Llama Anti-VP1 Recombinant Antibody (HPAB-0504-YJ-VHH) | ELISA, IF, Inhib, WB | Llama VHH |
| HPAB-0505-YJ-VHH | Llama Anti-VP1 Recombinant Antibody (HPAB-0505-YJ-VHH) | ELISA, IF, Inhib, WB | Llama VHH |
| HPAB-0506-YJ-VHH | Llama Anti-VP1 Recombinant Antibody (HPAB-0506-YJ-VHH) | ELISA, IF, Inhib, WB | Llama VHH |
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