Revision 5

#29002

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Orders:

877-616-CELL (2355)

orders@cellsignal.com

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
IHC-Bond, IHC-P

Reactivity:
H

Sensitivity:
Endogenous

Source/Isotype:
Mouse IgG2b kappa

UniProt ID:
#P15056

Entrez-Gene Id:
673

Product Usage Information

Application Dilution
IHC Leica Bond 1:100 - 1:400
Immunohistochemistry (Paraffin) 1:100 - 1:400

Storage

Supplied in a Tris-based buffer with 1% BSA and less than 0.1% sodium azide. Stable for 24 months when stored at 4°C. Do not aliquot the antibody.

Specificity/Sensitivity

B-Raf (V600E Mutant) (IHC600) Mouse mAb recognizes the V600E mutant of B-Raf protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a peptide covering the V600E mutation.

Background

A-Raf, B-Raf, and c-Raf (Raf-1) are the main effectors recruited by GTP-bound Ras to activate the MEK-MAP kinase pathway (1). Activation of c-Raf is the best understood and involves phosphorylation at multiple activating sites, including Ser338, Tyr341, Thr491, Ser494, Ser497, and Ser499 (2). p21-activated kinase (PAK) has been shown to phosphorylate c-Raf at Ser338, and the Src family phosphorylates Tyr341 to induce c-Raf activity (3,4). Ser338 of c-Raf corresponds to similar sites in A-Raf (Ser299) and B-Raf (Ser445), although this site is constitutively phosphorylated in B-Raf (5). Inhibitory 14-3-3 binding sites on c-Raf (Ser259 and Ser621) can be phosphorylated by Akt and AMPK, respectively (6,7). While A-Raf, B-Raf, and c-Raf are similar in sequence and function, differential regulation has been observed (8). Of particular interest, B-Raf contains three consensus Akt phosphorylation sites (Ser364, Ser428, and Thr439) and lacks a site equivalent to Tyr341 of c-Raf (8,9). Research studies have shown that the B-Raf mutation V600E results in elevated kinase activity and is commonly found in malignant melanoma (10). Six residues of c-Raf (Ser29, Ser43, Ser289, Ser296, Ser301, and Ser642) become hyperphosphorylated in a manner consistent with c-Raf inactivation. The hyperphosphorylation of these six sites is dependent on downstream MEK signaling and renders c-Raf unresponsive to subsequent activation events (11).

Species Reactivity

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

Applications Key

IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin)

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalStain 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.

Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma using B-Raf (V600E Mutant) (IHC600) Mouse mAb performed on the Leica® BOND™ Rx. Mutational status was confirmed by genomic sequencing.
Immunohistochemistry Image 1: B-Raf (V600E Mutant) (IHC600) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human colon carcinoma (left, B-Raf V600E positive) and human colon carcinoma (right, B-Raf V600E negative) using B-Raf (V600E Mutant) (IHC600) Mouse mAb. Mutational status was confirmed by genomic sequencing.
Immunohistochemistry Image 1: B-Raf (V600E Mutant) (IHC600) Mouse mAb
Immunohistochemical analysis of paraffin-embedded A-375 xenograft (left, B-Raf V600E positive) and MKN74 xenograft (right, wild-type B-Raf positive) using B-Raf (V600E Mutant) (IHC600) Mouse mAb.
Immunohistochemistry Image 2: B-Raf (V600E Mutant) (IHC600) Mouse mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded 293T cell pellet, untransfected (left), B-Raf wild-type transfected (middle), and B-Raf V600E mutant transfected (right), using B-Raf (V600E Mutant) (IHC600) Mouse mAb.
Immunohistochemistry Image 3: B-Raf (V600E Mutant) (IHC600) Mouse mAb
Immunohistochemical analysis of paraffin-embedded SK-MEL-28 cell pellet (left, positive) or OVMANA cell pellet (right, negative) using B-Raf (V600E Mutant) (IHC600) Mouse mAb.
Immunohistochemistry Image 4: B-Raf (V600E Mutant) (IHC600) Mouse mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.