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Products:Signal Transduction >> Cytoskeleton / ECM >> Extracellular Matrix >> Structures >> Focal Adhesions
Overview
Product name
Anti-VASP antibody
See all VASP products (14) ...
Description
Rabbit polyclonal to VASP
Tested applications
ICC/IF, Flow Cyt, WBmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Predicted to work with
Dog
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human VASP.
(Peptide available as ab32244.)
Positive control
This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; A431; HUVEC; A549; NIH 3T3; MEF1; PC12.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Signal Transduction >> Cytoskeleton / ECM >> Extracellular Matrix >> Structures >> Focal Adhesions
Applications
Show applications key
Our Abpromise guarantee covers the use of ab26650 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use a concentration of 5 µg/ml
Flow Cyt: 1/200((see Abreview))
WB: Use a concentration of 1 - 5 µg/ml.Detects a band of approximately 43 kDa (predicted molecular weight: 40 kDa).
Target
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
modifications
Major 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.
Target information above from: UniProt accessionP50552
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Vasodilator stimulated phosphoprotein antibody
- Vasodilator-stimulated phosphoprotein antibody
- VASP antibody
- VASP_HUMAN antibody
Database links
- Entrez Gene: 403936 Dog
- Entrez Gene: 7408 Human
- Entrez Gene: 22323 Mouse
- Entrez Gene: 361517 Rat
- Omim: 601703 Human
- SwissProt: P50551 Dog
- SwissProt: P50552 Human
- SwissProt: P70460 Mouse
see all
Anti-VASP antibody images:
Western blot - VASP antibody (ab26650)
All lanes : Anti-VASP antibody (ab26650) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 40 kDa
Observed band size : 43 kDa (why is the actual band size different from the predicted?)
Additional bands at : 49 kDa (possible cross reactivity).
Western blot - VASP antibody (ab26650)
All lanes : Anti-VASP antibody (ab26650) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 40 kDa
Observed band size : 43 kDa (why is the actual band size different from the predicted?)
Immunocytochemistry/ Immunofluorescence - VASP antibody (ab26650)
ICC/IF image of ab26650 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab26650, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Western blot - VASP antibody (ab26650)
All lanes : Anti-VASP antibody (ab26650) at 1 µg/ml
Lane 1 : HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate
Lane 2 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 40 kDa
Observed band size : 45,49 kDa (why is the actual band size different from the predicted?)
Additional bands at : 86 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 20 minutes
Flow Cytometry - VASP antibody (ab26650)
ab26650 detecting VASP in Human platelets by Flow Cytometry. Platelets were isolated spinning Platelet rich plasma on Histopaque. Cells were fixed in PFA and permeabilized in 0.1% Triton-X100 in 2% BSA for 30 minutes. The primary antibody was diluted 1/200 in 2% BSA in PBS and incubated with the sample for 18 hours at 4°C. An Alexa Fluor® 488-conjugated Chicken anti-Rabbit IgG (H+L) antibody, diluted 1/500 was used as the secondary antibody.
Abbreviations in the figure:
Plts: Platelets;
UP: Unpermeabilized;
US Unstained, Red Peak;
IgG Rb: IgG Rabbit (Isotype Control), Blue Peak;
VASP: VASP, Green peak.
This image is courtesy of Mahesh Shivananjappa.
References for Anti-VASP antibody (ab26650)
ab26650 has not yet been referenced specifically in any publications.
Publishing research using ab26650? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"