Catalog # DP4521

mDia3 (Ser-196))[conserved site], phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 130 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). mDia3 is activated by Cdc42 and regulates the attachment of microtubules to the kinetochore during mitosis. Aurora B kinase phosphorylates Ser-196 and Thr-882 in vitro, and phosphorylation of Ser-196 increases during mitosis in vivo. A multisite mutant mDia3 with nonphosphorylatable T66A, S196A, S880A and T882A leads to misalignment of chromosomes at the metaphase plate. Thus, phosphorylation may be an important regulator of mDia3 activity during mitosis.

References

Cheng, L. et al. (2011) Developmental Cell 20:342.
Lammers, M. et al. (2008) J Biol Chem. 283(50):35236.
Yasuda, S. et al. (2004) Nature. 428(6984):767.

Western blot analysis of mDia3 expression in human Jurkat cells treated with Calyculin A (100 nM) (lanes 1-4). The blots were treated with lambda phosphatase (lanes 2 & 4), then probed with rabbit polyclonal anti-mDia3 (C-terminus; DP4511) (lanes 1 & 2) and anti-phospho-mDia3 (Ser-196; DP4521) (lanes 3 & 4).

Phospho-mDia3 (Ser-196) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding Ser-196 in human mDia3 (Diap2). This peptide sequence is identical in rat and mouse mDia3, and the site is well conserved in human mDia1/Diap1 (Ser-171) and mDia2/Diap3 (Ser-202).

*For more information, see UniProt Accession Q17R91

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 phospho-mDia3 (Ser-196) peptide (without carrier). The antibody detects a 130 kDa* protein corresponding to the apparent molecular mass of mDia3 on SDS-PAGE immunoblots of human Jurkat treated with calyculin A. This reactivity is not observed after lambda phosphatase treatment. The antibody may also detect phosphorylated mDia1/Diap1 (Ser-171) and mDia2/Diap3 (Ser-202).

*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: