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Technical Data Sheet

AK6480

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α-Actinin 4 Phospho-Regulation
Antibody Sampler Kit
Price

$395

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
AP4241 Kojima, S. et al. (2014) Am J Nephrol. 39(1):36-45 WB: human kidney cells
AP4271 Kojima, S. et al. (2014) Am J Nephrol. 39(1):36-45 WB: human kidney cells
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
Kit Summary
The α-Actinin 4 phospho-regulation antibody sampler kit can be used to examine phosphorylation of α-Actinin 4 at Tyr-4. The kit includes monoclonal anti-α-Actinin and polyclonal anti-α-Actinin 4 antibodies to monitor total expression levels of α-Actinins. Secondary reagents are also included for primary antibody detection.
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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).

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Western blot analysis of α-actinin in human A431 cells (lanes 1 & 3) and rabbit spleen fibroblasts (lanes 2 & 4). The blots were probed with rabbit polyclonal anti-α-actinin 4 (a.a. 2-11) or mouse monoclonal anti-α-actinin (C-terminal region).

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Background
α-Actinins are widely expressed cytoskeletal proteins that cross-link actin filaments through antiparallel 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 inhibits actin binding and reduces actin-filament driven multinucleation in rat kidney cells. Thus, phosphorylation in α-actinins may be important for regulating actin binding and actin cytoskeletal remodeling.

Background References
Ylanne et al. (2001) Structure. 9:597.
Izaguirre, G. et al. (2001) J Biol Chem. 276(31):28676.
Shao, H. et al. (2010) J Biol Chem. 285(4):2591.
Buffer and Storage
Mouse monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.
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