Recombinant Anti-VASP antibody [EPR1337(2)] (ab109321)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1337(2)] to VASP
- Suitable for: WB, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-VASP antibody [EPR1337(2)]
See all VASP primary antibodies -
Description
Rabbit monoclonal [EPR1337(2)] to VASP -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, Jurkat, HEK293, 293T, HepG2, THP-1 and human platelet lysates. IP: HEK293 cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1337(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109321 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Detects a band of approximately 46 kDa (predicted molecular weight: 40 kDa).
For unpurified use at 1/10000. |
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IP |
1/70.
For unpurified use at 1/10 - 1/100. |
Notes |
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WB
1/1000. Detects a band of approximately 46 kDa (predicted molecular weight: 40 kDa). For unpurified use at 1/10000. |
IP
1/70. For unpurified use at 1/10 - 1/100. |
Target
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Function
Ena/VASP proteins are actin-associated proteins involved in a range of processes dependent on cytoskeleton remodeling and cell polarity such as axon guidance, lamellipodial and filopodial dynamics, platelet activation and cell migration. VASP promotes actin filament elongation. It protects the barbed end of growing actin filaments against capping and increases the rate of actin polymerization in the presence of capping protein. VASP stimulates actin filament elongation by promoting the transfer of profilin-bound actin monomers onto the barbed end of growing actin filaments. Plays a role in actin-based mobility of Listeria monocytogenes in host cells. Regulates actin dynamics in platelets and plays an important role in regulating platelet aggregation. -
Tissue specificity
Highly expressed in platelets. -
Sequence similarities
Belongs to the Ena/VASP family.
Contains 1 WH1 domain. -
Domain
The EVH2 domain is comprised of 3 regions. Block A is a thymosin-like domain required for G-actin binding. The KLKR motif within this block is essential for the G-actin binding and for actin polymerization. Block B is required for F-actin binding and subcellular location, and Block C for tetramerization.
The WH1 domain mediates interaction with XIRP1. -
Post-translational
modificationsMajor substrate for cAMP-dependent (PKA) and cGMP-dependent protein kinase (PKG) in platelets. The preferred site for PKA is Ser-157, the preferred site for PKG, Ser-239. In ADP-activated platelets, phosphorylation by PKA or PKG on Ser-157 leads to fibrinogen receptor inhibition. Phosphorylation on Thr-278 requires prior phosphorylation on Ser-157 and Ser-239. In response to phorbol ester (PMA) stimulation, phosphorylated by PKC/PRKCA. In response to thrombin, phosphorylated by both PKC and ROCK1. Phosphorylation at Thr-278 by AMPK does not require prior phosphorylation at Ser-157 or Ser-239. Phosphorylation modulates F-actin binding, actin filament elongation and platelet activation. Carbon monoxide (CO) promotes phosphorylation at Ser-157, while nitric oxide (NO) promotes phosphorylation at Ser-157, but also at Ser-239. Response to NO and CO is blunted in platelets from diabetic patients, and VASP is not phosphorylated efficiently at Ser-157 and Ser-239. -
Cellular localization
Cytoplasm. Cytoplasm > cytoskeleton. Cell junction > focal adhesion. Cell projection > lamellipodium membrane. Cell projection > filopodium membrane. Targeted to stress fibers and focal adhesions through interaction with a number of proteins including MRL family members. Localizes to the plasma membrane in protruding lamellipodia and filopodial tips. Stimulation by thrombin or PMA, also translocates VASP to focal adhesions. Localized along the sides of actin filaments throughout the peripheral cytoplasm under basal conditions. - Information by UniProt
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Database links
- Entrez Gene: 7408 Human
- Omim: 601703 Human
- SwissProt: P50552 Human
- Unigene: 515469 Human
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Alternative names
- Vasodilator stimulated phosphoprotein antibody
- Vasodilator-stimulated phosphoprotein antibody
- VASP antibody
- VASP_HUMAN antibody
Images
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All lanes : Anti-VASP antibody [EPR1337(2)] (ab109321) at 1/1000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : VASP knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 40 kDa
Observed band size: 46 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab109321 observed at 46 kDa. Red - loading control ab8245 observed at 36 kDa.
ab109321 Anti-VASP antibody [EPR1337(2)] was shown to specifically react with VASP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265892 (knockout cell lysate ab257792) was used. Wild-type and VASP knockout samples were subjected to SDS-PAGE. ab109321 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Anti-VASP antibody [EPR1337(2)] (ab109321) at 1/1000 dilution (purified) + HEK293 cell lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 46 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-VASP antibody [EPR1337(2)] (ab109321) at 1/1000 dilution (purified) + HepG2 cell lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 46 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-VASP antibody [EPR1337(2)] (ab109321) at 1/5000 dilution (purified) + THP-1 cell lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 46 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-VASP antibody [EPR1337(2)] (ab109321) at 1/10000 dilution (unpurified)
Lane 1 : 293T cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : Human platelet lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 40 kDa
Observed band size: 46 kDa why is the actual band size different from the predicted? -
ab109321 (purified) at 1/30 immunoprecipitating VASP in HEK293 cell lysate (Lane 1). Lane 2 - rabbit monoclonal IgG instead of ab109321 in HEK293 lysates. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab109321 has been referenced in 1 publication.
- Marsden AJ et al. Inhibition of Arginine Methylation Impairs Platelet Function. ACS Pharmacol Transl Sci 4:1567-1577 (2021). PubMed: 34661075