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Technical Data Sheet

AP4241

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α-Actinin 4 (Tyr-4), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
104 kDa

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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).

Application
Dilution


ELISA
1:2000


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


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