Mouse / IgG2b
Anti-phosphotyrosine antibody is a tyrosine phosphorylation specific antibody that is part of the Signal-Seeker™ product line. APY03 anti-phosphotyrosine mouse monoclonal antibody clone 27B10 has been tested in western blot, immunofluorescence, immunoprecipitation and ELISA with excellent specificity and low background with strong potent signals. APY03 phosphotyrosine antibody recognizes a wide range of tyrosine phosphorylated proteins in cells treated with kinase inhibitors, hydrogen peroxide (H2O2) and sodium vanadate (Figure 1).
APY03 is a mouse monoclonal antibody 27B10 that recognizes phosphorytated tyrosine residues. APY03 was raised against a proprietary mixture of phosphotyrosine peptides conjugated to KLH. APY03 can detect 10 ng of phosphotyrosine-labeled bovine serum albumin (see Certificate of Analysis [COA]). APY03 is purified by protein G affinity chromatography and is supplied as a lyophilized white powder. Each Lot of antibody is quality controlled to provide a high batch to batch consistency. The Lot specific µg per tube can be found in the Lot specific COA documents at www.cytoskeleton.com. APY03 shows high specificity to phosphotyrosine peptides and does not cross-react with phosphoserine/threonine peptides in a western blot or an ELISA assay (Figures 1 & 4).
Figure 1: Western Blot using Phosphotyrosine Antibody
APY03 is highly specific for p-Tyr and does not recognize p-Ser or p-Thr tagged proteins. To see the full Western blot protocol, see the product datasheet.
Figure 2: Immunoprecipitation using Phosphotyrosine Antibody
APY03 was able to enrich a wide range of tyrosine-phosphorylated proteins from NIH3T3 cells treated with H2O2-activated orthovanadate. No signal was detected with Protein G bead control without APY03. To see the full Immunoprecipitation protocol, see the product datasheet.
Human epidermoid carcinoma A431 cells, untreated (3A) or treated (3B) with EGF (100 ng/ml for 3 minutes), and NIH3T3, untreated (3C) or treated (3D) with H2O2-activated sodium orthovanadate (100 µM for 10 minutes), were stained as described in the method. Phosphotyrosine and nuclei were visualized in green fluorescence and blue DAPI staining, respectively. To see the full Immunofluorescence protocol, see the product datasheet.
Figure 4: ELISA using Phosphotyrosine Antibody
Data show that APY03 does not crossreact with phosphothreonine and phoophoserine modified BSA. To see the full ELISA protocol, see the product datasheet.
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Signal-Seeker™ Phosphotyrosine Detection Kit (Cat. # BK160)
Signal-Seeker™ Phosphotyrosine Affinity Beads (Cat.# APY03-beads)
Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)
For product Datasheets and MSDSs please click on the PDF links below.
Sample Size Datasheet (Cat. APY03-S):
Certificate of Analysis: Lot 011
|Ahler, Ethan et al.||A Combined Approach Reveals a Regulatory Mechanism Coupling Src's Kinase Activity, Localization, and Phosphotransferase-Independent Functions||Molecular Cell||2019||ISSN 1097-4164|
|Kline, Ashley et al.||The Misshapen kinase regulates the size and stability of the germline ring canals in the Drosophila egg chamber||Developmental Biology||2018||ISSN 1095-564X|
|Seki, Takahiro et al.||Ablation of endothelial VEG FR1 improves metabolic dysfunction by inducing adipose tissue browning||Journal of Experimental Medicine||2018||ISSN 1540-9538|
|Cheng, Larry C. et al.||Phosphopeptide enrichment coupled with label-free quantitative mass spectrometry to investigate the phosphoproteome in prostate cancer||Journal of Visualized Experiments||2018||ISSN 1940-087X|
|Horita, Henrick et al.||Utilizing a comprehensive immunoprecipitation enrichment system to identify an endogenous post-translational modification profile for target proteins||Journal of Visualized Experiments||2018||ISSN 1940-087X|
|Horita, Henrick et al.||Utilizing optimized tools to investigate PTM crosstalk: Identifying potential PTM crosstalk of acetylated mitochondrial proteins||Proteomes||2018||ISSN 2227-7382|
|Horita, Henrick et al.||A simple toolset to identify endogenous post-translational modifications for a target protein: A snapshot of the EGFR signaling pathway||Bioscience Reports||2017||ISSN 1573-4935|
|Horita, Henrick et al.||Identifying Regulatory Posttranslational Modifications of PD-L1: A Focus on Monoubiquitinaton||Neoplasia (United States)||2017||ISSN 1476-5586|
|Kaukonen, Riina et al.||Normal stroma suppresses cancer cell proliferation via mechanosensitive regulation of JMJD1a-mediated transcription||Nature Communications||2016||ISSN 2041-1723|
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