α-Actinin-4, pAb



ALX-210-356-C050	50 µg	588.00 USD

Product Specification

Concentration:	250µg/ml

Purity Detail:	Epitope-affinity purified.

Formulation:	Liquid. In PBS containing 1mg/ml BSA and 0.01% sodium azide.

Immunogen:	Synthetic peptide corresponding to aa 28-38 (M²⁸GDYMAQEDDW³⁸) of human and rat actinin-4 and to aa 29-39 of mouse actinin-4.

Source/Host:	From rabbit.

Specificity:	Recognizes human, mouse, rat, pig, and Chinese hamster actinin-4. Does not cross-react with actinin-1, -2 or -3.

Application:	Immunohistochemistry (frozen sections (1µg/ml), paraffin sections) Immunoprecipitation (25µg/ml) Western Blot (0.1-0.25µg/ml) The recommended dilutions refer to the analysis of mouse cells and tissues with intermediate levels of actinin-4 expression and must be viewed as approximate.

Short Term Storage:	+4°C

Long Term Storage:	-20°C

Handling:	Avoid freeze/thaw cycles.

Positive Control:	Human platelet protein (500µg) (Prod. No. ALX-840-007), supplied at 5mg/ml in SDS sample buffer (100mM sodium chloride, 73mM TRIS/HCl pH 6.7, 10mM DTT, 8mM EDTA, 5% glycerol, 2% SDS, 10µg/ml Bromophenol Blue sodium salt).Use 5µl (25µg) per lane for Western blotting of tricine gels (13% acrylamide) or 15% Laemmli gels.

Miscellaneous/General:	α-Actinin-4 is an actin-bundling protein of ~100kDa that is associated with cell motility, endocytosis, and cancer invasion. The α-actinin family comprises two non-muscle isoforms (α-actinin-1 and -4) and two skeletal muscle isoforms (α-actinin-2 and -3), with α-actinin-2 being also expressed in cardiac muscle. While α-actinin-4 is almost ubiquitously expressed, particularly high concentrations are found in glomeruli. On the subcellular level it is associated with actin stress fibers, but in certain cells it also localizes to the nucleus. Mutations in the α-actinin-4 gene cause an autosomal-dominant form of familial focal segmental glomerulosclerosis (FSGS), which is thought to result from a defect in glomerular podocyte function. A point mutation in the α-actinin-4 gene was found to generate an antigenic peptide that is recognized by autologous cytolytic T lymphocytes (CTL) on a human lung carcinoma. α-actinin-4 interacts with a variety of proteins, including the ring finger protein BERP, the PDZ-LIM protein CLP-36, the hemidesmosomal and cell-cell contact protein BP180, and the tight junction protein MAGI-1. Moreover, α-actinin-4 forms a ternary complex with Ca²/ Calmodulin-dependent protein kinase II and Densin-180, a protein of postsynaptic densities in CNS neurons. Ca²-dependent association of α-actinin-4 with E3KARP is required for Ca²-dependent inhibition of the Na/ Hexchanger 3 (NHE3).

Figure 1: Western blot of mouse kidney (lane 1) and brain (lane 2) using PAb to α-Actinin-4 (Prod. No. ALX-210-356).

Figure 2: Immunohistochemistry of actinin-4 distribution in rat kidney using PAb to α-Actinin-4 (Prod. No. ALX-210-356).

Figure 3: Isoform and species cross-reactivity in immunoblots determined with solid phase bound synthetic peptides.
Spot Sequence               Isoform               Species
1 QTNDYMQPEEDWD     actinin 1               human, mouse, rat, chicken
2 DEDEYMIQEEEWD       actinin 2               human, mouse
3 GGGEYMEQEEDWD     actinin 3            human, mouse, rat
4 SMGDYMAQEDDWD     actinin 4               human, mouse, rat
5 GMGDYMRPGGDWD    actinin 4 variant    rat
6 MDAWYSDEQMDGD     Control scrambled actinin 4 peptide

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Product Literature References

CART: an Hrs/actinin-4/BERP/myosin V protein complex required for efficient receptor recycling: Q. Yan, et al.; Mol. Biol. Cell 16, 2470 (2005), Abstract; Full Text

Podocyte involvement in human immune crescentic glomerulonephritis: J. Bariety, et al.; Kidney Int. 68, 1109 (2005), Abstract;

Expression of alpha-actinin-4 in acquired human nephrotic syndrome: a quantitative immunoelectron microscopy study: N.P. Goode, et al.; Nephrol. Dial. Transplant. 19, 844 (2004), Abstract; Full Text

General Literature References

Ca(2+)-dependent inhibition of Na+/H+ exchanger 3 (NHE3) requires an NHE3-E3KARP-alpha-actinin-4 complex for oligomerization and endocytosis: J.H. Kim, et al.; J. Biol. Chem. 277, 23714 (2002), Abstract;

Interaction of two actin-binding proteins, synaptopodin and alpha-actinin-4, with the tight junction protein MAGI-1: K.M. Patrie, et al.; J. Biol. Chem. 277, 30183 (2002), Abstract;

A point mutation in the alpha-actinin-4 gene generates an antigenic peptide recognized by autologous cytolytic T lymphocytes on a human lung carcinoma: H. Echchakir, et al.; Cancer Res. 61, 4078 (2001), Abstract;

Densin-180 forms a ternary complex with the (alpha)-subunit of Ca2+/calmodulin-dependent protein kinase II and (alpha)-actinin: R.S. Walikonis, et al.; J. Neurosci. 21, 423 (2001), Abstract;

Interactions of a hemidesmosome component and actinin family members: A.M. Gonzalez, et al.; J. Cell Sci. 114, 4197 (2001), Abstract;

Actinin-4 is preferentially involved in circular ruffling and macropinocytosis in mouse macrophages: analysis by fluorescence ratio imaging: N. Araki, et al.; J. Cell Sci. 113, 3329 (2000), Abstract;

BERP, a novel ring finger protein, binds to alpha-actinin-4: A.E. El-Husseini, et al.; BBRC 267, 906 (2000), Abstract;

CLP-36 PDZ-LIM protein associates with nonmuscle alpha-actinin-1 and alpha-actinin-4: T. Vallenius, et al.; J. Biol. Chem. 275, 11100 (2000), Abstract;

Mutations in ACTN4, encoding alpha-actinin-4, cause familial focal segmental glomerulosclerosis: J.M. Kaplan, et al.; Nat. Genet. 24, 251 (2000), Abstract;

Alpha-Actinins: D.R. Critchley & G. Flood; in: Guidebook to the Cytoskeletal and Motor Proteins, T. Kreis & R. Vale, Editors, Oxford University Press: Oxford p. 24 (1999),

Actinin-4, a novel actin-bundling protein associated with cell motility and cancer invasion: K. Honda, et al.; J. Cell Biol. 140, 1383 (1998), Abstract;