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Technical Data Sheet

AX4245

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Phospho-α-Actinin 4 (Tyr-4)
Blocking Peptide
Price
Size

$55
50 μg

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Application
Dilution


Blocking
1:1000


ELISA
50 ng/well



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.
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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.
Sequence
Phospho-α-actinin 4 (Tyr-4) synthetic peptide corresponds 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
Blocking peptide is supplied in 50µl phosphate-buffered saline and 0.05% sodium azide.
Store at –20°C. Stable for 1 year.
Specificity
The peptide is specifically recognized by anti-α-actinin 4 (Tyr-4) phospho-specific antibody (AP4241) in ELISA, and has been shown to block the reactivity of AP4241 during Western blot. In addition, the peptide is recommended for use in blocking AP4241 reactivity in immunocytochemistry.
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