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.
References
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-specific.
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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*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blotmobilities of known proteins with similar MW.
Product References:
AP4241 Kojima, S. et al. (2014) Am J Nephrol. 39(1):36-45 WB: human kidney cellsAP4271 Kojima, S. et al. (2014) Am J Nephrol. 39(1):36-45 WB: human kidney cellsRS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblastsRS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western BlotThis kit contains:
| CATALOG# | DESCRIPTION | SIZE | APPLICATIONS | SPECIES REACTIVITY | MW (kDa) |
AP4271 | α-Actinin 4 (a.a. 2-11) Rabbit pAb | 50 μl | WB, E | Hu, Rt, Ms | 104 |
AX4275 | α-Actinin 4 (a.a. 2-11) Peptide | 50 μg | AB, E | ||
AP4241 | α-Actinin 4 (Tyr-4), phospho-specific Rabbit pAb | 50 μl | WB, E | Hu, Rt, Ms | 104 |
AX4245 | phospho-α-Actinin 4 (Tyr-4) Peptide | 50 μg | AB, E | ||
RS3251 | Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb | 100 μl | WB, E, ICC, IHC | Rb |
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 polyclonal anti-α-Actinin 4 antibody to monitor total expression levels of α-Actinins, peptides for blocking experiments, and HRP-conjugated secondary reagent for primary antibody detection.
