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553009 Sigma-AldrichPhosphoDetect™ Anti-c-Raf (pTyr³⁴⁰, Tyr³⁴¹) Rabbit pAb

This PhosphoDetect™ Anti-c-Raf (pTyr³⁴⁰, Tyr³⁴¹) Rabbit pAb is validated for use in WB, ICC for the detection of c-Raf (pTyr³⁴⁰, Tyr³⁴¹).

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

SDS
CoA
References
Data Sheet

Recommended Products

Overview

Replacement Information

Key Spec Table

Species Reactivity	Host	Antibody Type
H, M, R	Rb	Polyclonal Antibody

Pricing & Availability

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Description
Overview	Recognizes c-Raf phosphorylated at Tyr³⁴⁰ and Tyr³⁴¹ in HEK293 cells overexpressing c-Raf.
Overview	Note: 1 T = 1 test. Sufficient for 10 Western miniblots.
Catalogue Number	553009
Brand Family	Calbiochem®
Application Data	Detection of human c-Raf, phosphorylated on Tyr³⁴⁰, by immunoblotting. Samples: Whole cell lysate from HEK293 cells overexpressing c-Raf left unstimulated (lane 1) or stimulated with active Src (lane 2-5). Primary antibody: PhosphoDetect™ Anti-c-Raf (pTyr^{340, Tyr}341) Rabbit pAb (Cat. No. 553009) (1:000) pre-incubated without peptide (lanes 1, 2), with non-phosphopeptide corresponding to the immunogen (lane 3), a generic phosphoserine containing peptide (lane 4), or phosphopeptide immunogen (lane 5). Detection: chemiluminescence.

References
References	Yan, F., and Polk, D.B. 2001. Cancer Res. 61, 963. Yip-Schneider, M.T., et al. 2000. Biochem. J. 351, 151. Mason, C.S. et al. 1999. EMBO J. 18, 2137. Bonner, T.I. et al. 1985. Mol. Cell. Biol. 5, 1400. Moelling, K. et al. 1984. Nature 312, 558. Rapp, U.R. et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4218.

Product Information
Form	Liquid
Formulation	In phosphate buffer (without Mg²⁺ and Ca²⁺), 1 mg/ml BSA, 50% glycerol, pH 7.3 (+/- 0.1).
Positive control	HEK293 cells over expressing c-Raf, phosphorylated in vitro by Src.
Preservative	≤0.1% sodium azide
Quality Level	MQ100

Applications
Key Applications	Immunoblotting (Western Blotting) Immunocytochemistry
Application Notes	Immunoblotting (1:1000) Immunocytochemistry (see comments)
Application Comments	The immunogen sequence is 100% conserved in mouse and rat, but cross-reactivity has not been tested. This antibody has also been reported to work for immunocytochemistry. Variables associated with assay conditions will dictate the proper working dilution.

Biological Information
Immunogen	a synthetic phosphopeptide corresponding to amino acids surrounding the Tyr³⁴⁰ and Tyr³⁴¹, phosphorylation sites of human c-Raf (Raf-1)
Immunogen	Human
Host	Rabbit
Isotype	IgG
Species Reactivity	Human Mouse Rat
Antibody Type	Polyclonal Antibody

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements

Storage and Shipping Information
Ship Code	Blue Ice Only
Toxicity	Standard Handling
Storage	-20°C
Avoid freeze/thaw	Avoid freeze/thaw
Do not freeze	Ok to freeze
Special Instructions	Following initial thaw, aliquot and freeze (-20°C).

Packaging Information

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
553009-10T	04055977193411

Documentation

PhosphoDetect™ Anti-c-Raf (pTyr³⁴⁰, Tyr³⁴¹) Rabbit pAb SDS

Title
Safety Data Sheet (SDS)

PhosphoDetect™ Anti-c-Raf (pTyr³⁴⁰, Tyr³⁴¹) Rabbit pAb Certificates of Analysis

Title	Lot Number
553009	Enter Lot Number

References

Reference overview
Yan, F., and Polk, D.B. 2001. Cancer Res. 61, 963. Yip-Schneider, M.T., et al. 2000. Biochem. J. 351, 151. Mason, C.S. et al. 1999. EMBO J. 18, 2137. Bonner, T.I. et al. 1985. Mol. Cell. Biol. 5, 1400. Moelling, K. et al. 1984. Nature 312, 558. Rapp, U.R. et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4218.

Data Sheet

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision	15-August-2007 RFH
Application	Immunoblotting (1:1000) Immunocytochemistry (see comments)
Application Data	Detection of human c-Raf, phosphorylated on Tyr³⁴⁰, by immunoblotting. Samples: Whole cell lysate from HEK293 cells overexpressing c-Raf left unstimulated (lane 1) or stimulated with active Src (lane 2-5). Primary antibody: PhosphoDetect™ Anti-c-Raf (pTyr^{340, Tyr}341) Rabbit pAb (Cat. No. 553009) (1:000) pre-incubated without peptide (lanes 1, 2), with non-phosphopeptide corresponding to the immunogen (lane 3), a generic phosphoserine containing peptide (lane 4), or phosphopeptide immunogen (lane 5). Detection: chemiluminescence.
Description	Rabbit polyclonal antibody adsorbed against non-phosphopeptide corresponding to the immunogen phosphorylation site, followed by immunoaffinity chromatography. Recognizes the ~74 KDa c-Raf protein phosphorylated on Tyr^340/341.
Background	The raf family of proto-oncogenes consists of three members: A-raf-1, B-raf, and c-raf-1 which show the greatest degree of homology in the carboxy-terminal half of the molecule. In the case of c-raf-1 this region has been shown to have protein (serine/threonine) kinase activity. C-raf was first identified in an oncogeneic form, v-raf, which is the oncogene of the transforming virus MSV. The raf-1 gene is the most studied of the three and encodes a protein with an apparent molecular mass of ~74 kDa. Raf is widely expressed and plays an important role in cell proliferation and differentiation. Raf relays signals related to the activation of growth factor receptor to the activation of MAPK and utlimately to the nucleus. Activation of raf itself is dependent on tyrosine (341) phosphorylation. Tyrosine phosphorylation is accomplished by Src tyrosine kinase.
Host	Rabbit
Immunogen species	Human
Immunogen	a synthetic phosphopeptide corresponding to amino acids surrounding the Tyr³⁴⁰ and Tyr³⁴¹, phosphorylation sites of human c-Raf (Raf-1)
Isotype	IgG
Species	human, mouse, rat
Positive control	HEK293 cells over expressing c-Raf, phosphorylated in vitro by Src.
Form	Liquid
Formulation	In phosphate buffer (without Mg²⁺ and Ca²⁺), 1 mg/ml BSA, 50% glycerol, pH 7.3 (+/- 0.1).
Preservative	≤0.1% sodium azide
Comments	The immunogen sequence is 100% conserved in mouse and rat, but cross-reactivity has not been tested. This antibody has also been reported to work for immunocytochemistry. Variables associated with assay conditions will dictate the proper working dilution.
Storage	Avoid freeze/thaw -20°C
Do Not Freeze	Ok to freeze
Special Instructions	Following initial thaw, aliquot and freeze (-20°C).
Toxicity	Standard Handling
References	Yan, F., and Polk, D.B. 2001. Cancer Res. 61, 963. Yip-Schneider, M.T., et al. 2000. Biochem. J. 351, 151. Mason, C.S. et al. 1999. EMBO J. 18, 2137. Bonner, T.I. et al. 1985. Mol. Cell. Biol. 5, 1400. Moelling, K. et al. 1984. Nature 312, 558. Rapp, U.R. et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4218.

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