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.

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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.


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

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.

Target

  • Alternative Names
  • Capsid protein VP1; vp1

For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

See other products for "VP1"

* Abbreviations
3D IHC3D Immunohistochemistry
ActivActivation
AgonistAgonist
ApopApoptosis
BABioassay
BIBioimaging
BlockBlocking
Cell ScreeningCell Screening
SeparationCell Separation
ChIPChromatin Immunoprecipitation
CMCDComplement Mediated Cell Depletion
CostimCostimulation
CytCytotoxicity
DepletionDepletion
DBDot Blot
EMElectron Microscopy
ELISAEnzyme-linked Immunosorbent Assay
ELISPOTEnzyme-linked Immunosorbent Spot
FCFlow Cytometry
FuncSFunctional Assay
GSGel Super Shift Assay
HAHemagglutination
IAImmunoassay
IBImmunoblotting
ICCImmunocytochemistry
IDImmunodiffusion
IFImmunofluorescence
IHCImmunohistochemistry
IHC-FrImmunohistochemistry-Frozen
IHC-PImmunohistochemistry-Paraffin
REImmunohistology - Resin Sections
IPImmunoprecipitation
IRMAImmunoradiometric Assay
SHIn situ hybridization
InhibInhibition
ICFCIntracellular Staining for Flow Cytometry
KO/KD-WBKnockout/Knockdown target confirmation by Western Blot
Live cell imagingLive cell imaging
CyTOF®Mass Cytometry
MeDIPMethylated DNA Immunoprecipitation
MultiplexMultiplex bead-based assay
NeutNeutralization
PPProtein Purification
PGProteogenomics
RIRadial Immunodiffusion
RIARadioimmunoassay
StimStimulation
SPRSurface Plasmon Resonance
TCTissue Culture
TBTurbidimetry
WBWestern Blot

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