Catalog # DK6630


mDia Phospho-Regulation

Antibody Sampler Kit

$395

Formins include several families of proteins that regulate actin cytoskeletal dynamics via two conserved formin homology domains, FH1 and FH2. Formins construct actin-based structures comprising linear, unbranched filaments that are used in stress fibers, actin cables, microspikes, and contractile rings. The diaphanous (Dia) formin subfamily includes mDia1 (Diap1), mDia2 (Diap3), and mDia3 (Diap2). mDia1 protein is activated by Rho leading to ROCK-dependent stress fiber formation and regulation of serum response factor-dependent transcription. mDia2 is activated by Rac or Cdc42, and it nucleates actin polymerization and stabilizes microtubules. Regulation of mDia2 activity may occur through ROCK phosphorylation of Thr-1061 and Ser-1070. 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.

References
Miki, T. et al. (2009) J Biol Chem. 284:5753.
Narumiya, S. et al. (2009) Cancer Metastasis Rev. 28:65.
Staus, D.P. et al. (2011) Biochem J. 439(1):57-65.
Cheng, L. et al. (2011) Developmental Cell 20:342
Rabbit polyclonal, affinity-purified antibodies are supplied in 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.



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

Product References:

DP3491 Li X, et al. (2017) Haematologica. 102(6):984. ICC: fetal liver cells
DP3491 Monzo, P. et al. (2016) Mol Biol Cell. 27(8):1246. WB: rat C6 glioma, shRNA knockdown
DP3491 Spence, EF et al. (2016) J Neurosci.36(37):9696. WB: mouse postnatal brain
DP3491 Belin, BJ et al. (2015) Elife. Aug. 19;4:e07735. WB: human HeLa, shRNA knockdown
DP4511 Vitiello, E. et. al(2014) Nat Commun. 18(5):5826 WB: HeLa, HCE
DP3491 Sandbo, N. et al. (2013) J Biol Chem 288(22):15466. WB : human lung fibroblasts

Western blot analysis of mDia2 expression in rat PC12 (lane 1), human A431 (lane 2), mouse brain (lane 3), and rabbit spleen fibroblasts (lane 4). The blots were probed with anti-mDia2 (C-terminal region) at 1:500.

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).



This kit contains:

CATALOG #DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
mDia1 (a.a. 66-77) Rabbit pAb50 μlWB, EHu, Rt, Ms140
mDia2 (C-terminal region) Rabbit pAb50 μlWB, EHu, Rt, Ms135
mDia2 (Thr-1061), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms, Ck, F135
mDia3 (C-terminus) Rabbit pAb50 μlWB, EHu, Rt, Ms130
mDia3 (Ser-196)[conserved site], phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms130
KIT SUMMARY

The mDia phospho-regulation antibody sampler kit can be used to detect the expression levels of mDia1, mDia2, and mDia3. In addition, the kit contains phospho-specific antibodies to detect phoshorylation of Thr-1061 on mDia2, and phosphorylation of Ser-196 in mDia3. The latter antibody detects a site that is well conserved in mDia1 (Ser-171) and mDia2 (Ser-202), so the antibody may be useful for detection of phosphorylation of these sites as well.