Revision 3
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

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

210

Source/Isotype:

Mouse IgG2a

UniProt ID:

#A9UF07

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

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

Bcr-Abl (b2a2 Junction Specific) (L99H4) Mouse mAb detects endogenous levels of Bcr-Abl (b2a2) fusion proteins. This antibody does not cross-react with the b3a2 isoform of Bcr-Abl.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the b2a2 junction site sequence of human Bcr-Abl.

Background

The Bcr gene was orginally identified by its presence in the chimeric Bcr-Abl oncogene (1). The amino-terminal region of Bcr contains an oligomerization domain, a serine/threonine kinase domain, and a region that binds SH2 domains. The middle of the protein has a PH domain and a region of sequence similarity to the guanine nucleotide exchange factors for the Rho family of GTP binding proteins. The carboxy-terminal region may be involved in a GTPase activating function for the small GTP-binding protein Rac (2,3). The function of wild type Bcr in cells remains unclear. PDGF receptor may use Bcr as a downstream signaling mediator (4). Research studies have shown that the Bcr-Abl fusion results in production of a constitutively active tyrosine kinase, which causes chronic myelogenous leukemia (CML) (5). Tyr177 of Bcr is phosphorylated in the Bcr-Abl fusion protein, which plays an important role in transforming the activity of Bcr-Abl (6). Phosphorylated Tyr177 provides a docking site for Gab2 and GRB2 (7,8).
The fusion protein encoded by Bcr-Abl varies in size, depending on the breakpoint in the BCR gene. Three breakpoint cluster regions have been characterized to date: major (M-bcr), minor (m-bcr) and micro (mu-bcr). The overwhelming majority of CML patients have a p210 Bcr-Abl gene (M-bcr), whose mRNA transcripts have a b3a2 and/or a b2a2 junction. The smallest of the fusion proteins, p190 Bcr-Abl, (m-bcr breakpoint) is principally associated with Ph-positive ALL. Rare cases of CML are due to a p190-type of Bcr-Abl gene and in these, the disease tends to have a prominent monocytic component, resembling CMML. CML resulting from a p230 Bcr-Abl gene (mu-bcr breakpoint) is also rare, and has been associated with the CNL variant and/or with marked thrombocytosis. Exceptional CML cases have been described with Bcr breakpoints outside the three defined cluster regions, or with unusual breakpoints in Abl (9).

  1. Groffen, J. et al. (1984) Cell 36, 93-99.
  2. Maru, Y. et al. (1991) Cell 67, 459-468.
  3. Che, W. et al. (2001) Circulation 104, 1399-1406.
  4. Abe, J. I. et al. (2001) Ann. N.Y. Acad. Sci. 947, 341-343.
  5. Voncken, J. W. et al. (1995) Cell 80, 719-728.
  6. He, Y. et al. (2002) Blood 99, 2957-2968.
  7. Sattler, M. et al. (2002) Cancer Cell 1, 479-492.
  8. Warmuth, M. et al. (1995) J. Biol. Chem. 272, 33260-33270.
  9. Melo, J.V. (1997) Baillieres Clin. Haematol 10, 203-22.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting

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 3
#3908

Bcr-Abl (b2a2 Junction Specific) (L99H4) Mouse mAb

Western Blotting Image 1: Bcr-Abl (b2a2 Junction Specific) (L99H4) Mouse mAb Expand Image
Western blot analysis of various cell lysates from cell lines expressing Bcr-Abl fusion proteins, using Bcr-Abl (b2a2 Junction Specific) (L99H4) Mouse mAb (upper) and c-Abl Antibody #2862 (lower).