Catalog # DP4481

mDia2 (Thr-1061), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms, Ck, F

MW 135 kDa

Formins include several families of proteins that regulate actin cytoskeletal dynamics via two conserved formin homology domains, FH1 and FH2. Through cooperation of FH1 and FH2, formins construct actin-based structures comprising linear, unbranched filaments that are used in stress fibers, actin cables, microspikes, and contractile rings. A subgroup of the formins is the diaphanous (Dia) family, which includes mDia1 (Diap1), mDia2 (Diap3), and mDia3 (Diap2). The mDia2 protein has been implicated in cell migration and cytokinesis. This Dia protein can nucleate actin polymerization and stabilize microtubules. mDia2 may also function in the nucleus as the protein continually shuttles between the cytoplasm and nucleus. Regulation of mDia2 activity may occur through ROCK phosphorylation of Thr-1061 and Ser-1070. This phosphorylation disrupts inhibitory interactions between the DAD and DID domains leading to enhanced mDia2 activation of RhoA.

References

Staus, D.P. et al. (2011) Biochem J. 439(1):57-65.
Miki, T. et al. (2009) J Biol Chem. 284:5753.
Bartolini, F. et al. (2008) J Cell Biol. 181:523.
Alberts, A.S. (2001) J Biol Chem. 276:2824.

Western blot analysis of COS-7 cells expressing flag-tagged mDia2 (lanes 1-3) or mDia2 mutant (T1061A) (lane 4). The cells were untreated (lane 1) or treated with calyculin A (lanes 2 & 3). The blot was treated with alkaline phosphatase (lane 2), then probed with anti-mDia2 (Thr-1061) (upper blot) or anti-flag antibody (lower blot). (Image provided by Dr. Christopher Mack in the Dept. of Pathology at UNC Chapel Hill).

Phospho-mDia2 (Thr-1061) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding Thr-1061 in mouse mDia2 (Diap3). This peptide sequence is identical in rat and has 2 amino acid differences to human mDia2. The sequence has low homology to other formins.

*For more information, see UniProt Accession Q9Z207

Rabbit polyclonal, affinity-purified antibody is supplied in 100μl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store 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.

This antibody was affinity purified using mDia2 (Thr-1061) peptide (without carrier). The antibody detects a 135 kDa* protein corresponding to mDia2 in immunoblots of human A7r5 and mouse C2C12 cells treated with calyculin A. The antibody detects transfected mDia2 after treatment with calyculin A, but this reactivity is removed after phosphatase treatment. In addition, the antibody does not detect mDia2 (T1061A) mutant.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: