A part of MilliporeSigma

05-321X | Anti-Phosphotyrosine Antibody, clone 4G10®

50 µg  
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      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      Vrt ICC, IHC, IP, WB M Purified Monoclonal Antibody
      Catalogue Number05-321X
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionAnti-Phosphotyrosine Antibody, clone 4G10®
      Background InformationSome of the tyrosine residues can be tagged with a phosphate group (phosphorylated) by protein kinases. (In its phosphorylated state, it is referred to as phosphotyrosine.). Tyrosine phosphorylation is considered as one of the key steps in signal transduction and regulation of enzymatic activity.
      The advent of anti-phosphotyrosine antibodies is one of significant events in signal transduction research. Before the availability of anti-phosphotyrosine antibodies, tyrosyl phospyhorylation of proteins and enzymes was investigated through hazardous and time-consuming radioactive experiments. Anti-phosphotyrosine antibodies are commonly used in western blots after the targeted proteins have been immunoprecipitated to measure the tyrosyl phosphorylation of the proteins. Anti-phosphotyrosine antibodies are also directly used on cell lysate to examine the overall change of tyrosine phosphorylation level in reponse to various treatments.
      Product Information
      • Untreated A-431 (negative control) and EGF treated A-431 (positive control) whole cell lysates.
        Included Positive Antigen Control: Catalog # 12-302, EGF-stimulated A431 cell lysate is provided as a free positive antigen control for western immunoblotting. Aliquot as desired, refreeze immediately, and store at -20°C. The lysate is stable for 6 months at -20°C. Before use, add 2.5 μL of 2-mercaptoethanol/100 μL of lysate and boil for 5 minutes to reduce the preparation. Load 20 μg of reduced lysate per lane for immunoblot analysis.
      PresentationPurified mouse monoclonal IgG2bκ in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide. Liquid at 2-8ºC.
      IgG2bκ mouse monoclonal antibody produced in vitro by mouse-mouse hybridoma 4G10® (FOX-NY [NS-1 derivative] myeloma x spleen cells).
      ApplicationAnti-Phosphotyrosine Antibody, clone 4G10 detects tyrosine phosphorylated proteins in all species. This unique monoclonal antibody is validated for use in IC, IH, IP, WB and is backed by hundreds of publications.
      Key Applications
      • Immunocytochemistry
      • Immunohistochemistry
      • Immunoprecipitation
      • Western Blotting
      Application NotesImmunoprecipitation:
      2-4 μg of a previous lot immunoprecipitated quantitatively the phosphotyrosine containing proteins in the lysate of a confluent culture (10 cm dish) of cells expressing an activated tyrosine kinase. To preserve phosphotyrosine, add: 0.2 mM sodium orthovanadate to the lysis buffer.
      Biological Information
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      SpecificityThis antibody recognizes tyrosine-phosphorylated proteins from all species.
      Species Reactivity
      • Vertebrates
      Antibody TypeMonoclonal Antibody
      • Phosphorylation
      Purification MethodProtein G-Sepharose™ Chromatography
      Molecular WeightDependent upon the molecular weight of the tyrosine phosphorylated protein being detected.
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely evaluated on EGF-treated human A431 carcinoma cells.

      Western Blot Analysis:
      0.5-2 μg/mL of this lot detected tyrosine-phosphorylated proteins in a modified RIPA lysate from EGF-treated human A431 carcinoma cells (Cohen, B., 1990; Druker, B. J.,1989; Kanakura, Y., 1991).
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at 2-8°C from date of receipt.

      For maximum recovery of the product, centrifuge the original vial prior to removing the cap. If the product has accidentally been frozen and thawed, spin it at 13,000 x g for 10 minutes at 2-8ºC. Save the supernatant for application.
      Packaging Information
      Material Size50 µg
      Transport Information
      Supplemental Information




      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      Anti-Phosphotyrosine, clone 4G10#174; - 2167780 2167780
      Anti-Phosphotyrosine, clone 4G10#174; - 2019714 2019714
      Anti-Phosphotyrosine, clone 4G10#174; - 2074899 2074899
      Anti-Phosphotyrosine, clone 4G10#174; - 2246076 2246076
      Anti-Phosphotyrosine, clone 4G10#174; - DAM1632064 DAM1632064
      Anti-Phosphotyrosine, clone 4G10#174; - DAM1676163 DAM1676163
      Anti-Phosphotyrosine, clone 4G10#174; - DAM1739149 DAM1739149
      Anti-Phosphotyrosine, clone 4G10#174; - DAM1767467 DAM1767467
      Anti-Phosphotyrosine, clone 4G10#174; - JBC1858521 JBC1858521
      Anti-Phosphotyrosine, clone 4G10#174; Monoclonal Antibody - DAM1813163 DAM1813163
      Anti-Phosphotyrosine, clone 4G10<sup>reg;</sup> - 27317 27317
      Anti-Phosphotyrosine, clone 4G10<sup>reg;</sup> - 29808 29808
      Anti-Phosphotyrosine, clone 4G10® - 2446847 2446847
      Anti-Phosphotyrosine, clone 4G10® - 2328344 2328344
      Anti-Phosphotyrosine, clone 4G10® -2506494 2506494
      Anti-Phosphotyrosine, clone 4G10® -2607273 2607273
      Anti-Phosphotyrosine, clone 4G10® -2683843 2683843
      Anti-Phosphotyrosine, clone 4G10® -2716680 2716680
      Anti-Phosphotyrosine, clone 4G10® -2759759 2759759
      Anti-Phosphotyrosine, clone 4G10® -2819645 2819645


      Reference overviewSpeciesPub Med ID
      Cross-linking of IgGs bound on circulating neutrophils leads to an activation of endothelial cells: possible role of rheumatoid factors in rheumatoid arthritis-associated vascular dysfunction.
      Rollet-Labelle, E; Vaillancourt, M; Marois, L; Newkirk, MM; Poubelle, PE; Naccache, PH
      Journal of inflammation (London, England) 10 27 2013

      Show Abstract
      23902799 23902799
      Centrosomal targeting of tyrosine kinase activity does not enhance oncogenicity in chronic myeloproliferative disorders.
      T Bochtler,M Kirsch,B Maier,J Bachmann,U Klingmüller,S Anderhub,A D Ho,A Krämer
      Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K 26 2012

      Show Abstract
      22015771 22015771
      ALK-activating homologous mutations in LTK induce cellular transformation.
      Roll, JD; Reuther, GW
      PloS one 7 e31733 2012

      Show Abstract
      22347506 22347506
      Wnt/β-catenin dependent cell proliferation underlies segmented lateral line morphogenesis.
      Aman, Andy, et al.
      Dev. Biol., 349: 470-82 (2011) 349 2011

      Show Abstract
      20974120 20974120
      Norrin attenuates protease-mediated death of transformed retinal ganglion cells.
      Lin, Song, et al.
      Mol. Vis., 15: 26-37 (2009) 2009

      Rat19137075 19137075
      An RNA interference screen identifies a novel regulator of target of rapamycin that mediates hypoxia suppression of translation in Drosophila S2 cells.
      Lee, SJ; Feldman, R; O'Farrell, PH
      Molecular biology of the cell 19 4051-61 2008

      Show Abstract Full Text Article
      18653470 18653470
      Polo kinase links the stress pathway to cell cycle control and tip growth in fission yeast.
      Janni Petersen, Iain M Hagan
      Nature 435 507-12 2005

      Show Abstract
      15917811 15917811

      Related Products & Applications

      Alternative Packsize

      Catalog Number Description  
      05-321 Anti-Phosphotyrosine Antibody, clone 4G10® Show Pricing & Availability
      05-321MG Anti-Phosphotyrosine Antibody, clone 4G10® Show Pricing & Availability

      Alternative Format

      Catalog Number Description  
      05-777 Anti-Phosphotyrosine Antibody, recombinant clone 4G10® Show Pricing & Availability
      16-101 Anti-Phosphotyrosine Antibody, clone 4G10®, agarose conjugate Show Pricing & Availability
      16-103 Anti-Phosphotyrosine Antibody, clone 4G10®, Biotin Conjugate Show Pricing & Availability
      16-104 Anti-Phosphotyrosine Antibody, clone 4G10®, FITC conjugate Show Pricing & Availability
      16-105 Anti-Phosphotyrosine Antibody, clone 4G10®, HRP conjugate Show Pricing & Availability
      16-199 Anti-Phosphotyrosine Antibody, recombinant clone 4G10®, agarose conjugate Show Pricing & Availability
      16-204 Anti-Phosphotyrosine Antibody, recombinant clone 4G10®, biotin conjugate Show Pricing & Availability
      16-205 Anti-Phosphotyrosine Antibody, recombinant clone 4G10®, FITC conjugate Show Pricing & Availability

      Included Positive Control

      Catalog Number Description  
      12-302 EGF-Stimulated A431 Cell Lysate Show Pricing & Availability

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      Life Science Research > Antibodies and Assays > Primary Antibodies