Catalog # AP4241

α-Actinin 4 (Tyr-4), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 104 kDa

α-Actinins are widely expressed cytoskeletal proteins that cross-link actin filaments through anti-parallel homodimers of the rod domains. Four α-actinin genes have been discovered in humans with α-actinin 1 and 4 being widely expressed in non-muscle cells. α-Actinins contain three conserved domains that include an N-terminal actin binding domain, four spectrin-like repeats in the central region, and a C-terminal calmodulin binding domain. α-Actinin cross-links the actin filament networks and associates the network to focal adhesion sites through binding of talin and vinculin. α-Actinin 1 is phosphorylated at Tyr-12 by FAK, while α-actinin 4 can be phosphorylated at Tyr-4 and Tyr-31 after EGF treatment. Tyr-4 and Tyr-31 phosphorylation inhibit actin binding and reduces actin-filament driven multi-nucleation in rat kidney cells. Thus, phosphorylation in α-actinins may be important for regulating actin binding and actin cytoskeletal remodeling.

References

Shao, H. et al. (2010) J Biol Chem. 285(4):2591.
Izaguirre, G. et al. (2001) J Biol Chem. 276(31):28676.
Ylanne et al. (2001) Structure. 9:597.

Western blot analysis of α-actinin 4 in A431 cells stimulated with pervanadate (1 mM) for 30 min (lanes 1,2,5.6) or after immunoprecipitation using α-actinin (C-terminal region) antibody in the absence (lanes 3 & 7) or presence of pervanadate-treated A431 cell lysate (lanes 4 & 8). Some lanes of the blot were treated with alkaline phosphatase (lanes 2 & 6). The blots were probed with anti-α-actinin (C-terminal region) or anti-α-actinin 4 (Tyr-4).

Phospho-α-actinin 4 (Tyr-4) synthetic peptide (coupled to carrier protein) corresponding to amino acids in the N-terminus of human α-actinin 4. This sequence is well conserved in rat and mouse α-actinin 4, but is not conserved in other α-actinins.

*For more information, see UniProt Accession O43707

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.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

The antibody detects a 100 kDa* protein corresponding to the molecular mass of α-actinin 4 on SDS-PAGE immunoblots of human A431 cells treated with pervanadate, but is not observed after akaline phosphatase treatment.

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

Product References:

Kojima, S. et al. (2014) Am J Nephrol. 39(1):36-45 (WB: human kidney cells)

This kit contains: