BackgroundImagesKit ComponentsBuffer & StorageReferencesReviews 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. ReferencesIzaguirre, 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. 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). Rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagent is supplied in the same buffer without azide, and the peptides are supplied in PBS and 0.05% sodium azide. Store all 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.*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#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)AP4271α-Actinin 4 (a.a. 2-11) Rabbit pAb50 μlWB, EHu, Rt, Ms104AX4275α-Actinin 4 (a.a. 2-11) Peptide50 μgAB, EAP4241 α-Actinin 4 (Tyr-4), phospho-specific Rabbit pAb 50 μlWB, EHu, Rt, Ms104AX4245phospho-α-Actinin 4 (Tyr-4) Peptide50 μgAB, ERS3251Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRbKIT SUMMARYThe 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.