Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)

Submit a review Submit a question
Western blot - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
  • Western blot - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
  • Flow Cytometry - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
  • Immunoprecipitation - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
  • Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1067Y] to VAV2 - BSA and Azide free
  • Suitable for: IP, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-VAV2 antibody [EP1067Y] - BSA and Azide free
    See all VAV2 primary antibodies

  • Description

    Rabbit monoclonal [EP1067Y] to VAV2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Human cervical carcinoma. WB: HeLa, HAP1, 293 cell lysates. Flow Cyt: HeLa cells. IP: HEK293 cell lysate.

  • General notes

    Ab239846 is the carrier-free version of ab52640. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab239846 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab239846 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
IHC-P
Human
IP
Human
WB
Human
Application Abreviews Notes
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 101 kDa (predicted molecular weight: 101 kDa).
Notes
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 101 kDa (predicted molecular weight: 101 kDa).

Target

  • Function

    Guanine nucleotide exchange factor for the Rho family of Ras-related GTPases. Plays an important role in angiogenesis. Its recruitement by phosphorylated EPHA2 is critical for EFNA1-induced RAC1 GTPase activation and vascular endothelial cell migration and assembly.

  • Tissue specificity

    Widely expressed.

  • Sequence similarities

    Contains 1 CH (calponin-homology) domain.
    Contains 1 DH (DBL-homology) domain.
    Contains 1 PH domain.
    Contains 1 phorbol-ester/DAG-type zinc finger.
    Contains 1 SH2 domain.
    Contains 2 SH3 domains.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Guanine nucleotide exchange factor VAV2 antibody
    • Oncogene VAV2 antibody
    • Protein vav 2 antibody
    • VAV 2 antibody
    • Vav 2 oncogene antibody
    • VAV-2 antibody
    • VAV2 antibody
    • VAV2 oncogene antibody
    • VAV2_HUMAN antibody
    see all

Images

  • Western blot - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    Western blot - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    All lanes : Anti-VAV2 antibody [EP1067Y] (ab52640) at 1/20000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : VAV2 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 101 kDa
    Observed band size: 100 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab52640).

      Lanes 1- 2: Merged signal (red and green). Green - ab52640 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab52640 was shown to react with VAV2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265318 (knockout cell lysate ab257794) was used. Wild-type HeLa and VAV2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52640 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 20000 dilution and a 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)

    Human cervical carcinoma stained with ab52640 at 1/50 - 1/100 dilution

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52640).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    Western blot - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    All lanes : Anti-VAV2 antibody [EP1067Y] (ab52640) at 1/20000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : VAV2 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 101 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab52640).

    Lanes 1 - 4: Merged signal (red and green). Green - ab52640 observed at 101 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab52640 was shown to specifically react with VAV2 in wild-type HAP1 cells as signal was lost in VAV2 knockout cells. Wild-type and VAV2 knockout samples were subjected to SDS-PAGE. Ab52640 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/20000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    Flow Cytometry - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)

    Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling VAV2 (red) with ab52640 at a 1/2500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52640).

  • Immunoprecipitation - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    Immunoprecipitation - Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    VAV2 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 10µg of Rabbit monoclonal to VAV2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52640.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 100kDa; VAV2.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52640).

  • Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)
    Anti-VAV2 antibody [EP1067Y] - BSA and Azide free (ab239846)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab239846? Please let us know so that we can cite the reference in this datasheet.

ab239846 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

There are currently no Customer reviews or Questions for ab239846.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com