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Technical Data Sheet

IP1291

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Integrin β4 (Tyr-1526), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
200 kDa

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Enlarge

Western blot analysis of A431 cells serum starved overnight (lanes 1, 3, & 5) and treated with pervanadate (1 mM) for 30 min (lanes 2, 4, & 6). The blots were probed with rabbit polyclonal anti-Integrin β4 (Tyr-1526) (lanes 1 & 2) and anti-Integrin β4 (Tyr-1494) (lanes 3 & 4) or with mouse monoclonal anti-Integrin β4 (lanes 5 & 6).

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Enlarge

Immunocytochemical labeling of integrin β4 in control (Top) and pervanadate-treated A431 cells (Bottom). The cells were labeled with mouse monoclonal anti-integrin β4 (Cytoplasmic region) (left) or rabbit polyclonals anti-integrin β4 (Tyr-1494) (middle) or anti-integrin β4 (Tyr-1526) (right), then the antibodies were detected using appropriate secondary antibodies conjugated to DyLight® 594.

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Application
Dilution


ELISA
1:2000


ICC
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Integrins are cell adhesion molecules that can mediate bidirectional transfer of signals across the plasma membrane. The cytoplasmic domains of integrin family members interact with components of the signal transduction apparatus within cells. Integrin α6β4receptors are found in basement membrane along with laminin-5. These receptors are expressed in epithelial, schwann, endothelial and some immune cells. The cytoplasmic domain of the Integrin β4 subunit recruits the adaptor protein Shc and is required for assembly of hemidesmosomes. Tyrosine phosphorylation of multiple sites within the cytoplasmic domain regulates these cellular events. In particular, tyrosine 1526 interacts with the phosphotyrosine binding domain of Shc and is required for Shc activation. In addition, tyrosine 1494 is required for integrin-mediated IRS-2 phosphorylation and activation of PI3-kinase. More importantly this site is critical for integrin α6β4 increases in carcinoma invasion.


Background References
Dans, M. et al. (2001). J Biol Chem. 276(2):1494-1502.
Shaw, L.M. (2001). Mol Cell Biol. 21(15):5082-5093.
Wang, L. et al. (2012) J Cell Physiol. 227(2):474.
Immunogen
Phospho-Integrin β4 (Tyr-1526) synthetic peptide (coupled to KLH) corresponding to amino acid residues around tyrosine 1526 of human Integrin β4. This peptide sequence is found in all three Integrin β4 isoforms and is highly conserved in rat and mouse Integrin β4.
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
This antibody was cross-adsorbed to phospho-tyrosine coupled to agarose and to unphosphorylated Integrin β4 (Tyr-1526) peptide before affinity purification using phospho-Integrin β4 (Tyr-1526) peptide (without carrier). The purified antibody detects a 200kDa* protein corresponding to the molecular mass of Integrin β4 on SDS-PAGE immunoblots of A431 and src-transformed Hct116 cells treated with pervanadate, but not in control cells.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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