Phospho-VASP (Ser157) Antibody - #AF3337

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Product: Phospho-VASP (Ser157) Antibody
Catalog: AF3337
Description: Rabbit polyclonal antibody to Phospho-VASP (Ser157)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Sheep, Dog
Mol.Wt.: 46kDa; 40kD(Calculated).
Uniprot: P50552
RRID: AB_2834752

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Related Downloads
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(88%), Bovine(83%), Sheep(83%), Dog(88%)
Clonality:
Polyclonal
Specificity:
Phospho-VASP (Ser157) Antibody detects endogenous levels of VASP only when phosphorylated at Serine 157.
RRID:
AB_2834752
Cite Format: Affinity Biosciences Cat# AF3337, RRID:AB_2834752.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Vasodilator stimulated phosphoprotein; Vasodilator-stimulated phosphoprotein; VASP; VASP_HUMAN;

Immunogens

Immunogen:
Uniprot:

P50552(VASP_HUMAN) >>Visit HPA database.

Gene(ID):
VASP(7408)
Expression:
P50552 VASP_HUMAN:

Highly expressed in platelets.

Description:
Vasodilator-stimulated phosphoprotein (VASP) is a member of the Ena-VASP protein family. Ena-VASP family members contain an EHV1 N-terminal domain that binds proteins containing E/DFPPPPXD/E motifs and targets Ena-VASP proteins to focal adhesions.
Sequence:
MSETVICSSRATVMLYDDGNKRWLPAGTGPQAFSRVQIYHNPTANSFRVVGRKMQPDQQVVINCAIVRGVKYNQATPNFHQWRDARQVWGLNFGSKEDAAQFAAGMASALEALEGGGPPPPPALPTWSVPNGPSPEEVEQQKRQQPGPSEHIERRVSNAGGPPAPPAGGPPPPPGPPPPPGPPPPPGLPPSGVPAAAHGAGGGPPPAPPLPAAQGPGGGGAGAPGLAAAIAGAKLRKVSKQEEASGGPTAPKAESGRSGGGGLMEEMNAMLARRRKATQVGEKTPKDESANQEEPEARVPAQSESVRRPWEKNSTTLPRMKSSSSVTTSETQPCTPSSSDYSDLQRVKQELLEEVKKELQKVKEEIIEAFVQELRKRGSP

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
88
Dog
88
Bovine
83
Sheep
83
Rabbit
63
Horse
0
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P50552 As Substrate

Site PTM Type Enzyme
S2 Acetylation
S2 Phosphorylation
S8 Phosphorylation
Y16 Phosphorylation
Y39 Phosphorylation Q9UM73 (ALK)
T43 Phosphorylation
S46 Phosphorylation
R68 Methylation
K71 Ubiquitination
S134 Phosphorylation
S157 Phosphorylation P17252 (PRKCA) , P17612 (PRKACA) , Q15139 (PRKD1) , Q13976 (PRKG1) , Q9BZL6 (PRKD2)
S239 Phosphorylation Q13976 (PRKG1) , Q13237 (PRKG2) , P17612 (PRKACA) , Q9BZL6 (PRKD2)
T249 Phosphorylation
K252 Acetylation
S258 Phosphorylation
T278 Phosphorylation Q15418 (RPS6KA1) , P54646 (PRKAA2) , P17612 (PRKACA) , Q13131 (PRKAA1) , Q13976 (PRKG1)
K283 Acetylation
T284 Phosphorylation
K286 Sumoylation
S289 Phosphorylation
S305 Phosphorylation
K312 Ubiquitination
S314 Phosphorylation
T315 Phosphorylation
T316 Phosphorylation
K321 Methylation
K321 Ubiquitination
S322 Phosphorylation P54646 (PRKAA2) , Q9BZL6 (PRKD2) , Q15139 (PRKD1)
S323 Phosphorylation
S324 Phosphorylation
S325 Phosphorylation
T327 Phosphorylation
T328 Phosphorylation
T331 Phosphorylation
T335 Phosphorylation
S337 Phosphorylation
Y341 Phosphorylation
K348 Ubiquitination
K363 Ubiquitination

Research Backgrounds

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.

PTMs:

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/PRKG1, 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 at Ser-157 by PKA is required for localization to the tight junctions in epithelial cells. Phosphorylation modulates F-actin binding, actin filament elongation and platelet activation. Phosphorylation at Ser-322 by AMPK also alters actin filament binding. 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.

Subcellular Location:

Cytoplasm. Cytoplasm>Cytoskeleton. Cell junction>Focal adhesion. Cell junction>Tight junction. Cell projection>Lamellipodium membrane. Cell projection>Filopodium membrane.
Note: 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. In pre-apoptotic cells, colocalizes with MEFV in large specks (pyroptosomes).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Highly expressed in platelets.

Subunit Structure:

Homotetramer. Interacts with PFN1, PFN2, LPP, ACTN1 and ACTG1. Interacts, via the EVH1 domain, with the Pro-rich regions of ZYX. This interaction is important for targeting to focal adhesions and the formation of actin-rich structures at the apical surface of cells. Interacts, via the EVH1 domain, with the Pro-rich domain of Listeria monocytogenes actA. Interacts with APBB1IP. Interacts, via the Pro-rich domain, with the C-terminal SH3 domain of DNMBP (By similarity). Interacts weakly with MEFV.

Family&Domains:

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.

Belongs to the Ena/VASP family.

Research Fields

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (View pathway)

· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Platelet activation.   (View pathway)

· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis.   (View pathway)

· Organismal Systems > Immune system > Leukocyte transendothelial migration.   (View pathway)

References

1). p47phox deficiency impairs platelet function and protects mice against arterial and venous thrombosis. Redox Biology (PubMed: 32422541) [IF=11.4]

Application: WB    Species: Human    Sample: platelets

Fig. 4. ROS generation and phosphorylation of VASP, ERK1/2, p38 MAPK, ERK5, JNK, AKT and c-PLA2. (A) Western blot analysis of the expression of NOX2, p67phox, NOX1, NOXO1 and Rac in WT and p47phox-/- platelets. (B) ROS generation in platelets after stimulation with CRP (2 μg/ml) or thrombin (0.5 U/ml) was expressed as mean fluorescent intensity (MFI) (mean ± SE, n = 6) (Student t-test). (C) The phosphorylation level of VASP, ERK1/2, p38, ERK5 and JNK in CRPstimulated platelets was detected by western blot and (D) quantified as a ratio relative to the total level (mean ± SD, n = 3) (Two-way ANOVA). (E) The phosphorylation level of AKT and c-PLA2 was also detected and (F) quantified (mean ± SD, n = 3) (Two-way ANOVA). *p < 0.05; **p < 0.01; ***p < 0.001.
2). Inhibition of Hypoxia-Inducible Factor Prolyl-Hydroxylase Modulates Platelet Function. THROMBOSIS AND HAEMOSTASIS (PubMed: 35477177) [IF=6.7]
3). Matrine Impairs Platelet Function and Thrombosis and Inhibits ROS Production. Frontiers in Pharmacology (PubMed: 34366869) [IF=5.6]

Application: WB    Species: Human    Sample:

FIGURE 6 ROS generation, phosphorylation of ERK1/2, p38, AKT and VASP and cGMP level. After matrine treatment, platelet ROS generation was measured using H2DCF-DA by flow cytometry (A). Matrine-treated platelets were stimulated with 5 μg/ml CRP or 1 U/ml thrombin for 5 min to measure the phosphorylation level of ERK1/2, p38, AKT (B and C) or VASP (D) by western blot (mean ± SD, n = 3), cGMP level by ELISA (mean ± SE, n = 3) (E). Compared with 0, * p < 0.05; **p < 0.01; ***p < 0.001.

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