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566807 Sigma-AldrichPhosphoDetect™ Anti-Shc/p66 (pSer³⁶) Mouse mAb (6E10)

This PhosphoDetect™ Anti-Shc/p66 (pSer³⁶) Mouse mAb (6E10) is validated for use in ELISA, Immunoblotting for the detection of Shc/p66 (pSer³⁶).

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

CoA
References
Data Sheet
Citations

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Overview

Replacement Information

Key Spec Table

Species Reactivity	Host	Antibody Type
H, M	M	Monoclonal Antibody

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Description
Overview	Recognizes the ~66 kDa form of Shc phosphorylated at Ser³⁶. Does not cross-react with the non-phosphorylated form of Shc/p66 or with unrelated phosphorylation sites.
Catalogue Number	566807
Brand Family	Calbiochem®
Application Data	Detection of human Shc/p66, phosphorylated on Ser³⁶, by immunoprecipitation. Samples: Timecourse of HeLa cells stimulated with EGF. Primary antibody: PhosphoDetect™ Anti-Shc/p66 (pSer³⁶) Mouse mAb (6E10) (Cat. No. 566807) (1 µg/ml). Detection: chemiluminescence.

References
References	Foschi, M., et al. 2001. J. Biol. Chem. 276, 26640. Gu, H., et al. 2000. Mol. Cell. Biol. 20, 7109. Migliaccio, E., et al. 1997. EMBO J. 16, 706. Okada, S., et al. 1997. J. Biol. Chem. 272, 28042. Pelicci, G., et al.1992. Cell 70, 93.

Product Information
Form	Lyophilized
Formulation	100 µg antibody lyophilized from 2X PBS, PEG, sucrose and 125 µl control protein lyophilized from 2X PBS.
Preservative	≤0.1% sodium azide (antibody only)
Quality Level	MQ100

Applications
Key Applications	Enzyme-Linked Immunosorbent Assay Immunoblotting (Western Blotting)
Application Notes	ELISA (0.05 µg/ml) Immunoblotting (0.1-1 µg/ml)
Application Comments	For immunoblotting using the p66/Shc positive control, use 15 µl per lane for colorimetric or chemiluminescent detection. Variables associated with assay conditions will dictate the proper working dilution.

Biological Information
Immunogen	a synthetic phosphopeptide corresponding to amino acids surrounding the Ser³⁶ phosphorylation site of human Shc/p66
Clone	6E10
Host	Mouse
Isotype	IgG₁
Species Reactivity	Human Mouse
Antibody Type	Monoclonal Antibody

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements

Storage and Shipping Information
Ship Code	Ambient Temperature Only
Toxicity	Standard Handling
Storage	-20°C
Do not freeze	Ok to freeze
Special Instructions	Reconstitute the antibody with 1 ml dH₂O for at least 15 min at room temperature until completely dissolved. Following reconstitution, aliquot and freeze (-20°C). Reconstitute the positive control with 125 µl H₂O until completely dissolved. After complete solubilization, add 125 µl SDS-PAGE sample buffer and incubate at 90°C for 5 min. Following reconstitution, aliquot and freeze (-20°C). Avoid freeze/thaw cycles of solutions.

Packaging Information

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
566807-1SETCN	04055977267082

Documentation

PhosphoDetect™ Anti-Shc/p66 (pSer³⁶) Mouse mAb (6E10) Certificates of Analysis

Title	Lot Number
566807	Enter Lot Number

References

Reference overview
Foschi, M., et al. 2001. J. Biol. Chem. 276, 26640. Gu, H., et al. 2000. Mol. Cell. Biol. 20, 7109. Migliaccio, E., et al. 1997. EMBO J. 16, 706. Okada, S., et al. 1997. J. Biol. Chem. 272, 28042. Pelicci, G., et al.1992. Cell 70, 93.

Citations

Title

Sun, L., et al. 2010. Am J Physiol Renal Physiol. 299, F1014.

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	ELISA (0.05 µg/ml) Immunoblotting (0.1-1 µg/ml)
Application Data	Detection of human Shc/p66, phosphorylated on Ser³⁶, by immunoprecipitation. Samples: Timecourse of HeLa cells stimulated with EGF. Primary antibody: PhosphoDetect™ Anti-Shc/p66 (pSer³⁶) Mouse mAb (6E10) (Cat. No. 566807) (1 µg/ml). Detection: chemiluminescence.
Description	Immunoaffinity purified mouse monoclonal antibody. Recognizes the Shc/p66 protein phosphorylated on Ser³⁶. Supplied with a positive control consisting of Shc/p66 from EGF-stimulated HepG2 cells immunoprecipitated using anti-Shc non-phospho-specific antibody.
Background	Mammalian cells can express three alternatively spliced isoforms of the shc adaptor protein: Shc/p46, Shc/p52 and Shc/p66. Shc/p46 and Shc/p52 are found in every cell type, whereas Shc/p66 expression varies in different tissues. Shc proteins are phosphorylated by receptor tyrosine kinases and interact with phosphotyrosine residues on these receptors via the shc PTB and/or SH2 domains. The major tyrosine phosphorylation sites are tyrosines 239/240 as well as tyrosine 317. The Shc/p66 protein contains an additional N-terminal protein domain that is phosphorylated at serine 36. It is suggested that only the serine-phosphorylated Shc/p66 can interact with GRB2. Moreover, expression of Shc/p66 accelerates the inactivation of MAP kinases after EGF stimulation, whereas Shc/p46 and Shc/p52 enhance MAP kinase activity.
Host	Mouse
Immunogen	a synthetic phosphopeptide corresponding to amino acids surrounding the Ser³⁶ phosphorylation site of human Shc/p66
Clone	6E10
Isotype	IgG₁
Species	human, mouse
Form	Lyophilized
Formulation	100 µg antibody lyophilized from 2X PBS, PEG, sucrose and 125 µl control protein lyophilized from 2X PBS.
Preservative	≤0.1% sodium azide (antibody only)
Comments	For immunoblotting using the p66/Shc positive control, use 15 µl per lane for colorimetric or chemiluminescent detection. Variables associated with assay conditions will dictate the proper working dilution.
Storage	-20°C
Do Not Freeze	Ok to freeze
Special Instructions	Reconstitute the antibody with 1 ml dH₂O for at least 15 min at room temperature until completely dissolved. Following reconstitution, aliquot and freeze (-20°C). Reconstitute the positive control with 125 µl H₂O until completely dissolved. After complete solubilization, add 125 µl SDS-PAGE sample buffer and incubate at 90°C for 5 min. Following reconstitution, aliquot and freeze (-20°C). Avoid freeze/thaw cycles of solutions.
Toxicity	Standard Handling
References	Foschi, M., et al. 2001. J. Biol. Chem. 276, 26640. Gu, H., et al. 2000. Mol. Cell. Biol. 20, 7109. Migliaccio, E., et al. 1997. EMBO J. 16, 706. Okada, S., et al. 1997. J. Biol. Chem. 272, 28042. Pelicci, G., et al.1992. Cell 70, 93.
Citation	Sun, L., et al. 2010. Am J Physiol Renal Physiol. 299, F1014.

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