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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human VAV1 (phospho Y174). The exact sequence is proprietary.
This product is a recombinant rabbit monoclonal antibody.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
Our Abpromise guarantee covers the use of ab76225 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/250000 - 1/500000. Detects a band of approximately 101 kDa (predicted molecular weight: 101 kDa).|
|Flow Cyt||Use at an assay dependent concentration.|
|ICC/IF||1/100 - 1/500.|
Immunocytochemistry/Immunofluorescence analysis of Untreated Jurkat cells and Pervanadate (1mM,30min) treated Jurkat cells labelling VAV1 (phospho Y174) with purified ab76225 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100, and counterstained with ab7291 anti-Tubulin (mouse mAb) 1:1000 (1ug/ml). An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Negative Control 1: Rabbit primary antibody and anti-mouse secondary antibody(ab150120)
Flow Cytometry analysis of Jurkat (human acute T cell leukemia) untreated/treated with 1mM pervanadate for 30min cells labeling VAV1 with unpurified ab76225 at 1/200 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. Untreated control (Green).
Dot blot analysis of VAV1 (pY174) peptide (Lane 1) and VAV1 non-phospho peptide (Lane 2) labelling VAV1 (phospho Y174) with ab76225 at a dilution of 1/1000. A Peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.