VASP Recombinant Rabbit Monoclonal Antibody [SY02-16]
Catalog# ET1607-6
VASP Recombinant Rabbit Monoclonal Antibody [SY02-16]
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WB
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IF-Cell
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IHC-P
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Human
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unconjugated
Overview
Product Name
VASP Recombinant Rabbit Monoclonal Antibody [SY02-16]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human VASP aa 51-100 / 380.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P
Molecular Weight
Predicted band size: 40 kDa
Positive Control
HeLa cell lysate, HT-29 cell lysate, HeLa, human lymph nodes tissue.
Conjugation
unconjugated
Clone Number
SY02-16
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IF-Cell
-
1:100
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IHC-P
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1:200
Target
Function
The Wiskott-Aldrich syndrome (WAS) is characterized by thrombocytopenia, eczema, defects in cell-mediated and humoral immunity, and a propensity for lymphoproliferative diseases. The syndrome is the result of a mutation in the gene encoding a proline-rich protein termed WASP. WASP has been identified as a downstream effector of Cdc42 and has been implicated in Actin polymer-ization and cytoskeletal organization. A distantly related protein, VASP (vaso-dilator-stimulated phosphoprotein), is involved in the maintenance of cytoarchitecture by interacting with Actin-like filaments. VASP shares a limited degree of homology with the amino-terminus of WASP, which is frequently mutated in WAS patients. An established substrate of cAMP and cGMP de-pendent kinases, VASP is phosphorylated on a regulatory Serine residue 157 and localizes to focal adhesions, microfilaments and highly active regions of the plasma membrane. VASP is highly expressed in human platelets and, like WASP, may play a role in cytoskeletal organization.
Background References
1. Bernusso, VA. et al. 2015. Imatinib restores VASP activity and its interaction with Zyxin in BCR-ABL leukemic cells. Biochimica et biophysica acta. 1853: 388-95.
2. Lee, NP. et al. 2004. Zyxin, axin, and Wiskott-Aldrich syndrome protein are adaptors that link the cadherin/catenin protein complex to the cytoskeleton at adherens junctions in the seminiferous epithelium of the rat testis. J Androl. 25(2): 200-215.
Sequence Similarity
Belongs to the Ena/VASP family.
Tissue Specificity
Highly expressed in platelets.
Post-translational Modification
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, Cell junction, Cell projection.
UNIPROT
Synonyms
Vasodilator stimulated phosphoprotein antibody
Vasodilator-stimulated phosphoprotein antibody
VASP antibody
VASP_HUMAN antibody
Images
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Western blot analysis of VASP on different lysates with Rabbit anti-VASP antibody (ET1607-6) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HT-29 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 40 kDa
Observed band size: 45 kDa
Exposure time: Lane 1: 17 seconds; Lane 2: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1607-6) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of VASP on different lysates with Rabbit anti-VASP antibody (ET1607-6) at 1/5,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-VASP KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 40 kDa
Observed band size: 45 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1607-6) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling VASP with Rabbit anti-VASP antibody (ET1607-6) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-VASP antibody (ET1607-6) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-VASP antibody (ET1607-6) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1607-6) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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