Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

120

Source/Isotype:

Rabbit IgG

UniProt ID:

#P35611

Entrez-Gene Id:

118

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-α-Adducin (Ser12) (E5X8Y) Rabbit mAb recognizes endogenous levels of α-adducin protein only when phosphorylated at Ser12. In some cell lines, this antibody cross-reacts with a 150 kDa band of unknown origin. This band appears to be upregulated in mitosis.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser12 of human α-adducin protein.

Background

The adducins (ADD) are cytoskeleton-associated proteins that help cap the ends of actin filaments, promote association between spectrin and actin, and participate in synapse assembly. The three closely related genes ADD1, ADD2, and ADD3 encode the α-adducin, β-adducin, and γ-adducin proteins (1). Research studies indicate that β-adducin is found at high levels in brain and hematopoietic tissues, whereas both α-adducin and γ-adducin are ubiquitously expressed (2). Adducin protein function is regulated by phosphorylation at a number of sites. Both PKA and PKC can phosphorylate α-adducin at Ser726 and β-adducin at Ser713, which inhibits calmodulin binding and adducin activity (3-5). Additionally, PKA (but not PKC) can phosphorylate β-adducin at Ser408, Ser436, and Ser481, which negatively affects spectrin-actin interactions (3). Phosphorylation of α-adducin at Thr445 and Thr480 by Rho-kinase regulates cell motility and membrane ruffling (6). Finally, CDK-1 phosphorylation of α-adducin at Ser12 and Ser355 during mitosis leads to association of α-adducin with the mitotic spindle, suggesting that α-adducin may play a role in mitotic regulation (7). Because α-adducin plays a role in regulating renal sodium reabsorption, it is not surprising that a number of studies show a relationship between ADD1 genetic polymorphisms and the development of hypertension (8-10).

  1. Matsuoka, Y. et al. (2000) Cell Mol Life Sci 57, 884-95.
  2. Joshi, R. et al. (1991) J Cell Biol 115, 665-75.
  3. Matsuoka, Y. et al. (1996) J Biol Chem 271, 25157-66.
  4. Chen, C.L. et al. (2007) J Cell Sci 120, 1157-67.
  5. Naydenov, N.G. and Ivanov, A.I. (2010) Mol Biol Cell 21, 3506-17.
  6. Fukata, Y. et al. (1999) J Cell Biol 145, 347-61.
  7. Chan, P.C. et al. (2014) J Cell Biol 204, 19-28.
  8. Kalita, J. et al. (2013) Neurol Res 35, 429-34.
  9. Kundu, A. and Anand, A. (2013) Cell Biochem Biophys 65, 13-9.
  10. Watanabe, Y. et al. (2010) Hypertens Res 33, 129-34.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#84214

Phospho-α-Adducin (Ser12) (E5X8Y) Rabbit mAb

Western Blotting Image 1: Phospho-α-Adducin (Ser12) (E5X8Y) Rabbit mAb Expand Image
Western blot analysis of extracts from HT-29 cells treated with thymidine (2 mM, 17 hr), released into fresh media for 8 hours, and treated again with thymidine (2 mM, 17 hr) (double thymidine block) followed by cell lysis (thymidine/thymidine) or released into media containing 100 ng/mL Nocodazole #2190 for 24 hours (thymidine/thymidine/nocodazole), using Phospho-α-Adducin (Ser12) (E5X8Y) Rabbit mAb (upper), α-Adducin (D7T7R) Rabbit mAb #70174 (middle) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: Phospho-α-Adducin (Ser12) (E5X8Y) Rabbit mAb Expand Image
Immunoprecipitation of phospho-α-adducin (Ser12) protein from HT-29 cell extracts treated with Nocodozole #2190 (100 ng/mL, 16 hr) to enrich cells in mitosis. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-α-Adducin (Ser12) (E5X8Y) Rabbit mAb. Western blot analysis was performed using

Phospho-α-Adducin (Ser12) (E5X8Y) Rabbit mAb.