Revision 1

#4880Store at -20C

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, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

130

SOURCE:

Rabbit

UniProt ID:

#Q8N163

Entrez-Gene Id:

57805

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:25
Immunofluorescence (Immunocytochemistry) 1:400

Storage

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

Specificity / Sensitivity

Phospho-DBC1 (Thr454) Antibody detects endogenous levels of DBC1 only when phosphorylated on Thr454.

Species Reactivity:

Human

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to Thr454 of the human DBC1 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Deleted in breast cancer gene 1 protein (DBC1) was originally identified by its localization to a region of chromosome 8p21 that is homozygously deleted in breast cancer (1). DBC1 is a large, nuclear protein with multiple functions in cell survival. It binds directly to the estrogen receptor α (ERα) hormone-binding domain in a ligand-independent manner and may be a key determinant of ligand-independent ERα expression and survival in human breast cancer cells (2). DBC1 can promote p53-mediated apoptosis by binding to and inhibiting the deacetylase activity of SirT1, resulting in increased p53 acetylation levels and activity (3). DBC1 may be an important regulator of heterochromatin formation as it binds SUV39H1 and inhibits its histone methyltransferase activity (4). Caspase-dependent processing activates the pro-apoptotic activity of DBC1 during Tumor Necrosis Factor-α (TNF-α)-mediated cell death signaling (5). This processing of DBC1 in response to TNF-α is an early event in the onset of apoptosis and results in relocalization of DBC1 to the cytoplasm. Overexpression of the processed, cytoplasmic form of DBC1 results in mitochondrial clustering and matrix condensation and sensitizes cells to TNF-α-mediated apoptosis.
The threonine residue at 454 of DBC1 is phosphorylated in an ATM/ATR-dependent manner in response to DNA damage (6,7). Phospho-DBC1 (Thr454) Antibody is directed at a site that was identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for modification site discovery. Phosphorylation at Thr454 was discovered using an ATM/ATR substrate antibody and was shown to be induced by UV treatment. Please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org for more information.

  1. Hamaguchi, M. et al. (2002) Proc Natl Acad Sci USA 99, 13647-52.
  2. Trauernicht, A.M. et al. (2007) Mol Endocrinol 21, 1526-36.
  3. Zhao, W. et al. (2008) Nature 451, 587-90.
  4. Li, Z. et al. (2009) J Biol Chem 284, 10361-6.
  5. Sundararajan, R. et al. (2005) Oncogene 24, 4908-20.
  6. Stokes, M.P. et al. (2007) Proc Natl Acad Sci USA 104, 19855-60.
  7. Beausoleil, S.A. et al. (2004) Proc Natl Acad Sci USA 101, 12130-5.

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 IF-IC: Immunofluorescence (Immunocytochemistry)

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.
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
#4880

Phospho-DBC1 (Thr454) Antibody

Western Blotting Image 1: Phospho-DBC1 (Thr454) Antibody Expand Image
Western blot analysis of extracts from 293 and HeLa cell lines, untreated or UV-treated (100 J/m2 and 4 hour recovery), using Phospho-DBC1 (Thr454) Antibody (upper) or β-actin Antibody #4967 (lower).
No image available
Immunofluorescence Image 1: Phospho-DBC1 (Thr454) Antibody Expand Image
Confocal immunofluorescent analysis of HeLa cells, untreated (left) and UV-treated (right), using Phospho-DBC1 (Thr454) Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).