VASP [p Ser156] Antibody (JE45-91)

Western Blot: VASP [p Ser156] Antibody (JE45-91) [NBP3-32815] - Western blot analysis of VASP on different lysates with Rabbit anti-VASP antibody (NBP3-32815) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate Lane 3: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, then the membrane treated with pp for 1 hour Lysates/proteins at 20 ug/Lane. Predicted band size: 40 kDa Observed band size: 50 kDa Exposure time: 3 minutes; ECL; 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 (NBP3-32815) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: VASP [p Ser156] Antibody (JE45-91) [NBP3-32815] - Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-VASP antibody (NBP3-32815) at 1/1,000 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 (NBP3-32815) at 1/1,000 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.

Immunocytochemistry/ Immunofluorescence: VASP [p Ser156] Antibody (JE45-91) [NBP3-32815] - Immunocytochemistry analysis of A431 cells treated with or without 10μM forskolin for 1 hour labeling VASP with Rabbit anti-VASP antibody (NBP3-32815) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 (NBP3-32815) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF, IHC
• Clone: JE45-91
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

VASP [p Ser156] Antibody (JE45-91) Summary

• Description: Novus Biologicals Rabbit VASP [p Ser156] Antibody (JE45-91) (NBP3-32815) is a recombinant monoclonal antibody validated for use in IHC, WB and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Synthetic phosphopeptide corresponding to residues surrounding Ser156 of human VASP. (Uniprot: P50552)
• Modification: p Ser156
• Localization: Cytoplasm, cytoskeleton, Cell junction, focal adhesion, tight junction, Cell projection, lamellipodium membrane, filopodium membrane.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: VASP
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:1000
  • Western Blot 1:1000
• Theoretical MW: 40 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for VASP [p Ser156] Antibody (JE45-91)

• vasodilator-stimulated phosphoprotein
• VASP

Background

Vasodilator-stimulated phosphoprotein (VASP) is a member of the Ena-VASP protein family. VASP is associated with filamentous actin formation and likely plays a widespread role in cell adhesion and motility. VASP may also be involved in the intracellular signaling pathways that regulate integrin-extracellular matrix interactions. VASP is regulated by the cyclic nucleotide-dependent kinases PKA and PKG. VASP shares a limited degree of homology with the amino terminus of WASP, which is frequently mutated in Wiskott-Aldrich syndrome (WAS) patients. An established substrate of cAMP and cGMP dependent 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: VASP