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).
Bulk Order Anti-mDia3 (Ser-196)) [conserved site], Phosphospecific Antibody
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.
Antigen Affinity Purified
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).
Human, Mouse, Rat
Storage at -20°C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20°C.
PBS + 1 mg/ml BSA, 0.05% NaN3 and 50% glycerol
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).
Western blots performed on each lot.
For research use only. Not intended for therapeutic or diagnostic use. Use of all products is subject to our terms and conditions, which can be viewed on our website.
After date of receipt, stable for at least 1 year at -20°C.