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Western blotting, suggested working dilution(s): Use 1-2 µg per ml antibody dilution buffer per mini-gel. Do not use dilution or blocking buffers containing milk as they may interfere with antibody binding to proteins of interest. Dilution and blocking buffers containing 4% bovine serum albumin are recommended for use with this antibody. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
Application Notes:
Additional reported applications (for the relevant formats) include: immunoprecipitation1,2, Western blotting1,2, immunofluorescence microscopy3.
Hela cell extract was resolved by electrophoresis, transferred to nitrocellulose, and probed with monoclonal anti-phosphotyrosine antibody (clone PY-20). Lane 1, serum-starved Hela cells; Lane 2, serum-starved Hela cells following serum addition for 4 hrs. Lane 2 shows an upregulation of tyrosine phosphorylated proteins after serum addition. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system.
Phosphorylation is a common modification of proteins that can result in alterations in protein function, protein-protein association, cellular localization, and protein-half life. Phosphorylation can occur on threonine, serine, and tyrosine residues. The PY20 monoclonal antibody recognizes phosphorylated tyrosine residues in all species tested (human, mouse, rat, dog, chicken, and frog). The PY20 antibody has been shown to be useful for flow cytometry, immunoprecipitation, Western blotting, and immunofluorescence staining.
Distribution:
Phospho-Specific
Function:
Phosphorylation of specific tyrosine residues, signal transduction, cell cycle progression, oncogenic transformation
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This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
Biotin anti-Phosphotyrosine
Hela cell extract was resolved by electrophoresis, transferred to nitrocellulose, and probed with anti-phosphotyrosine antibody (clone PY-20). Lane 1, serum-starved Hela cells; Lane 2, serum-starved Hela cells following serum addition for 4 hrs. Lane 2 shows an upregulation of tyrosine phosphorylated proteins after serum addition. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system.
Purified anti-Phosphotyrosine
Hela cell extract was resolved by electrophoresis, transferred to nitrocellulose, and probed with monoclonal anti-phosphotyrosine antibody (clone PY-20). Lane 1, serum-starved Hela cells; Lane 2, serum-starved Hela cells following serum addition for 4 hrs. Lane 2 shows an upregulation of tyrosine phosphorylated proteins after serum addition. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system.
