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Clone
G104
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Immunogen
Phosphotyrosine, alanine and glyceine in a 1:1:1 ratio polymerized in the presence of keyhole limpet hemocyanin with 1-ethyl-3-(3'-dimentrylaminopropyl) carbodiimide
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Antibody s full description
Mouse Anti- Phosphotyrosine Monoclonal IgG1 Antibody, Clone: G104: ATTO 594
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Antibody s category
Monoclonal Antibodies
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Antibody s other name
PhosphoTyrosine (pY) Antibody, PhosphoTyrosine (pY) Antibody
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Raised in
Mouse
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Antibody s target
Phosphotyrosine
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Primary research fields
Cell Signaling, Post-translational Modifications, Phosphorylation
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Brandname
None
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Antibodies applications
WB, IHC, ICC/IF, IP
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Antibody s reactivity
Species Independent
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Antibody s dilutions
WB (1:1000), IHC (1:100); optimal dilutions for assays should be determined by the user.
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Purity
Protein G Purified
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Antibody buffer for storage
PBS pH7.4, 50% glycerol, 0.09% sodium azide
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Antibody s concentration
1 mg/ml
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Antibody s specificity
Reacts with phosphotyrosine, and detects the presence of phosphotyrosine in both un-stimulated and stimulated cell lysates. Does not cross-react with phosphoserine or phosphothreonine.
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Storage recommendations
-20ºC
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Shipping recommendations
Blue Ice or 4ºC
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Antibody certificate of analysis
1 µg/ml of SMC-174 was sufficient for detection of phosphorylated tyrosine residues in 10 µg of rat tissue lysate by colorimetric immunoblot analysis using Goat anti-rat IgG:HRP as the secondary antibody.
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Antibody in cell
See included datasheet or contact our support service
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Tissue specificity
See included datasheet or contact our support service
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Scientific context
Protein phosphorylation is an important posttranslational modification that serves many key functions to regulate a protein’s activity, localization, and protein-protein interactions. Phosphorylation is catalyzed by various specific protein kinases, which involves removing a phosphate group from ATP and covalently attaching it to to a recipient protein that acts as a substrate. Most kinases act on both serine and threonine; others act on tyrosine, and a number (dual specificity kinases) act on all three. Because phosphorylation can occur at multiple sites on any given protein, it can therefore change the function or localization of that protein at any time (3). Changing the function of these proteins has been linked to a number of diseases, including cancer, diabetes, heart disease, inflammation and neurological disorders (4-6). In particular, the phosphorylation of tyrosine is considered one of the key steps in signal transduction and regulation of enzymatic activity (7). Phosphotyrosine can be detected through specific antibodies, and are helpful in facilitating the identification of tyrosine kinase substrates (8).
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Bibliography
1. Garton A.J., Flint A.J., and Tonks N.K. (1996) Mol and Cell Bio 16(11): 6408-6418. 2. Garton A.J., and Tonks N.K. (1999) J Bio Chem. 274(6): 3811-3818. 3. Goto H. et al. (2005) Nature Cell Biology 8: 180-187. 4. Blume-Jensen P. and Hunter T. (2001) Nature 411: 355-365. 5. Downward J. (2001) Nature 411: 759-762. 6. Pawson T. and Saxton T.M. (1999) Cell 97: 675-678. 7. Frackelton A.R. Jr., Ross A.H., and Eisen H.N. (1983) Mol Cell Biol. 3: 1343-1352. 8. Ross A.H., Baltimore D., and Eisen H.N. (1981) Nature 294: 654-656. 9. Tiganis T., Kemp B.E., and Tonks N.K. (1999) J. Bio Chem. 274(39): 27768-27775.
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Released date
1-Jan-2010
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Tested applications
To be tested
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Tested reactivity
To be tested
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NCBI number
Refer to NCBI
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Gene number
Refer to GenBank
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Protein number
Refer to Swiss-Prot
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Antibody s datasheet
Contact our support service
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Representative figure link
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Representative figure legend
Immunohistochemistry analysis using Mouse Anti-Phosphotyrosine Monoclonal Antibody, Clone G104 (SMC-174). Tissue: backskin. Species: Mouse. Fixation: Bouin's Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-Phosphotyrosine Monoclonal Antibody (SMC-174) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Stratum granulosum staining in the epidermis. Some dermal staining. Western Blot analysis of Human A431 cell lysates showing detection of Phosphotyrosine protein using Mouse Anti-Phosphotyrosine Monoclonal Antibody, Clone G104 (SMC-174). Load: 15 µg. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Phosphotyrosine Monoclonal Antibody (SMC-174) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT. Left: normal, right: EGF treated. Mouse Anti-Phosphotyrosine Antibody [G104] used in Immunohistochemistry (IHC) on Mouse backskin (SMC-174) Mouse Anti-Phosphotyrosine Antibody [G104] used in Western Blot (WB) on Human A431 cell lysates (SMC-174)
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Warning information
Non-hazardous
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Country of production
Canada
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Total weight kg
1.4
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Net weight g
0.1
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Stock availability
In Stock
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Description
This antibody needs to be stored at + 4°C in a fridge short term in a concentrated dilution. Freeze thaw will destroy a percentage in every cycle and should be avoided.
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About
Monoclonals of this antigen are available in different clones. Each murine monoclonal anibody has his own affinity specific for the clone. Mouse monoclonal antibodies are purified protein A or G and can be conjugated to FITC for flow cytometry or FACS and can be of different isotypes.
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Test
Mouse or mice from the Mus musculus species are used for production of mouse monoclonal antibodies or mabs and as research model for humans in your lab. Mouse are mature after 40 days for females and 55 days for males. The female mice are pregnant only 20 days and can give birth to 10 litters of 6-8 mice a year. Transgenic, knock-out, congenic and inbread strains are known for C57BL/6, A/J, BALB/c, SCID while the CD-1 is outbred as strain.
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Latin name
Mus musculus