antibody

Phosphotyrosine (HRP)

P4078-28

10Blots

monoclonal

mouse

HRP

4G10

Phosphotyrosine (HRP)

mouse

Mab

HRP

Antibodies

10Blots

4G10

IgG2b,k

Protein tyrosine phosphorylation has been the subject of intense investigation for over two decades. Tyrosine phosphorylation is catalyzed by members of the tyrosine kinase superfamily. To date, approximately 90 members of this superfamily have been identified in the human genome. Levels of phosphorylation of tyrosine residues within cellular proteins are observed to increase in response to growth factors, cytokines, insulin, extracellular matrix components, and other stimuli. Tyrosine phosphorylation has several effects, including the regulation of enzyme activity, as well as the localization of enzyme activity through the generation of docking sites for proteins bearing SH2 domains. Applications: Suitable for use in Western Blot and ELISA. Also, for use in applications in which an HRP conjugate is advantageous, such as Western Blot and Immunoprecipitation. Other applications not tested. Recommended Dilutions: Western Blot: 0.5-2ug/ml, with optimal performance at 2ug/ml (10ul per 10ml blocking buffer). Detects tyrosine-phosphorylated proteins in RIPA lysates from EGF-treated human A431 carcinoma cells. EGF-stimulated cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Phosphotyrosine, HRP conjugate (1ug/ml). Proteins were visualized using a chemiluminescence detection system. Optimal dilutions to be determined by the researcher. Positive Control: Pervanadate-treated human A431 cell extracts, EGF-treated human A431 cells. Storage and Stability: Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Note: Sodium azide is a potent inhibitor of peroxidase and should not be added to HRP conjugates. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Phosphotyramine-KLH.

Recognizes tyrosine-phosphorylated proteins from all species.

Purified by Protein G affinity chromatography.

Supplied as a liquid in PBS, 0.01% merthiolate. Sufficient for 10 immunoblots, if used at the optimum concentration of 2ug/ml in 10ml blocking buffer. Labeled with horseradish peroxidase (HRP)

~2mg/ml

Product Type: Mab Isotype: IgG2b,k Clone No: 4G10 Host: mouse Concentration: ~2mg/ml Form: Supplied as a liquid in PBS, 0.01% merthiolate. Sufficient for 10 immunoblots, if used at the optimum concentration of 2ug/ml in 10ml blocking buffer. Labeled with horseradish peroxidase (HRP) Purity: Purified by Protein G affinity chromatography. Immunogen: Phosphotyramine-KLH. Specificity: Recognizes tyrosine-phosphorylated proteins from all species. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

E IP WB

-20°C

1. McMahon, K.A., et al., Biochem J. 358: 431-436, 2001. 2. Blakesley, V.A., et al., J. Biol. Chem. 273: 18411-18422, 1998. 3. Elchebly, M., et al., Science 283: 1544-1548, 1999. 4. Field, K.A., et al., J. Biol. Chem. 272: 4276-4280, 1997. 5. Kracht, M. et al., Biochem J. 302:897-905, 1994. 6. Liossis, S.N., et al., J. Clin. Invest. 101: 1448-1457, 1998. 7. Cohen, B., et al., Proc. Natl. Acad. Sci. USA. 87: 4458-4462, 1990. 8. Druker, B.J., et al., New Eng. J. Med. 321: 1383-1391, 1989. 9. Kanakura, Y., et al., J. Biol. Chem. 266: 490-495, 1991.

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