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

05-321
100 µg  
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      Overview

      Replacement Information
      Replacement Information05-321 is a recommended replacement for MAB3090

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      AICC, IHC, IP, WBMPurifiedMonoclonal Antibody
      Description
      Catalogue Number05-321
      ReplacesMAB3090
      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.
      References
      Product Information
      FormatPurified
      Control
      • 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.
      Presentation0.1M Tris-Glycine, 0.15M NaCl, 0.05% Sodium Azide, pH 7.4. Liquid at 2-8°C.
      IgG2bκ mouse monocolonal antibody produced in vitro by mouse-mouse hybridoma 4G10® (FOX-NY [NS-1 derivative] myeloma x spleen cells).
      Applications
      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
      ImmunogenPhosphotyramine-KLH
      Clone4G10®
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityThis antibody recognizes tyrosine-phosphorylated proteins from all species.
      IsotypeIgG2bκ
      Species Reactivity
      • All
      Antibody TypeMonoclonal Antibody
      Modifications
      • Phosphorylation
      Purification MethodProtein G-Sepharose™ Chromatography
      Molecular WeightDependent upon the molecular weight of the tyrosine phosphorylated protein being detected.
      Physicochemical Information
      Dimensions
      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.

      NOTE: DO NOT FREEZE.
      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 Size100 µg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      Anti-Phosphotyrosine, clone 4G10#174; (mouse monoclonal IgG2bk) - DAM1423768 DAM1423768
      Anti-Phosphotyrosine, clone 4G10#174; - 2138006 2138006
      Anti-Phosphotyrosine, clone 4G10#174; - 2013168 2013168
      Anti-Phosphotyrosine, clone 4G10#174; - 2023919 2023919
      Anti-Phosphotyrosine, clone 4G10#174; - 2041109 2041109
      Anti-Phosphotyrosine, clone 4G10#174; - 2202535 2202535
      Anti-Phosphotyrosine, clone 4G10#174; - 2268815 2268815
      Anti-Phosphotyrosine, clone 4G10#174; - DAM1423768 DAM1423768
      Anti-Phosphotyrosine, clone 4G10#174; - DAM1503375 DAM1503375
      Anti-Phosphotyrosine, clone 4G10#174; - DAM1682114 DAM1682114

      References

      Reference overviewApplicationSpeciesPub Med ID
      Activity-regulated trafficking of the palmitoyl-acyl transferase DHHC5.
      Brigidi, GS; Santyr, B; Shimell, J; Jovellar, B; Bamji, SX
      Nature communications  6  8200  2015

      Show Abstract
      26334723 26334723
      Molecular characterization of WDCP, a novel fusion partner for the anaplastic lymphoma tyrosine kinase ALK.
      Yokoyama, N; Miller, WT
      Biomedical reports  3  9-13  2015

      Show Abstract
      25469238 25469238
      In vivo imaging and characterization of actin microridges.
      Lam, PY; Mangos, S; Green, JM; Reiser, J; Huttenlocher, A
      PloS one  10  e0115639  2015

      Show Abstract
      25629723 25629723
      P-TEFb, the super elongation complex and mediator regulate a subset of non-paused genes during early Drosophila embryo development.
      Dahlberg, O; Shilkova, O; Tang, M; Holmqvist, PH; Mannervik, M
      PLoS genetics  11  e1004971  2015

      Show Abstract
      25679530 25679530
      The tyrosine phosphatase SHP-1 regulates hypoxia inducible factor-1α (HIF-1α) protein levels in endothelial cells under hypoxia.
      Alig, SK; Stampnik, Y; Pircher, J; Rotter, R; Gaitzsch, E; Ribeiro, A; Wörnle, M; Krötz, F; Mannell, H
      PloS one  10  e0121113  2015

      Show Abstract
      25799543 25799543
      Phospho-tyrosine dependent protein-protein interaction network.
      Grossmann, A; Benlasfer, N; Birth, P; Hegele, A; Wachsmuth, F; Apelt, L; Stelzl, U
      Molecular systems biology  11  794  2015

      Show Abstract
      25814554 25814554
      HSP90 inhibitor AUY922 induces cell death by disruption of the Bcr-Abl, Jak2 and HSP90 signaling network complex in leukemia cells.
      Tao, W; Chakraborty, SN; Leng, X; Ma, H; Arlinghaus, RB
      Genes & cancer  6  19-29  2015

      Show Abstract
      25821558 25821558
      Novel agents that downregulate EGFR, HER2, and HER3 in parallel.
      Ferreira, RB; Law, ME; Jahn, SC; Davis, BJ; Heldermon, CD; Reinhard, M; Castellano, RK; Law, BK
      Oncotarget  6  10445-59  2015

      Show Abstract
      25865227 25865227
      ATP synthase promotes germ cell differentiation independent of oxidative phosphorylation.
      Teixeira, FK; Sanchez, CG; Hurd, TR; Seifert, JR; Czech, B; Preall, JB; Hannon, GJ; Lehmann, R
      Nature cell biology  17  689-96  2015

      Show Abstract
      25915123 25915123
      Trans-Activation between EphA and FGFR Regulates Self-Renewal and Differentiation of Mouse Embryonic Neural Stem/Progenitor Cells via Differential Activation of FRS2α.
      Sawada, T; Arai, D; Jing, X; Furushima, K; Chen, Q; Kawakami, K; Yokote, H; Miyajima, M; Sakaguchi, K
      PloS one  10  e0128826  2015

      Show Abstract
      26024354 26024354

      Brochure

      Title
      Western Blotting Tools

      FAQ

      QuestionAnswer
      Is Anti-Phosphotyrosine temperature sensitive?Anti-Phosphotyrosine is temperature sensitive and it must be stored at 4°C.
      Is Anti-Phosphotyrosine pH sensitive?Anti-Phosphotyrosine is extremely sensitive to acidic conditions. If exposed to a pH less than 3.0, the antibody loses most of its viability.
      What is tyramine?Tyramine is a normal substance in the body that helps support blood pressure; it is also found in certain foods. Researchers generated a phosphorylated form of Tyramine i.e., phosphotyramine.
      What is the difference between Anti-Phosphotyrosine, recombinant clone 4G10® (catalog #05-777) and Anti-Phosphotyrosine, clone 4G10® (catalog #05-321)?Anti-Phosphotyrosine, recombinant clone 4G10® (catalog #05-777) and Anti-Phosphotyrosine, clone 4G10® (catalog #05-321) are similar in performance. If you are interested in using Anti-Phosphotyrosine over a long period of time, you may want to use the Anti-Phosphotyrosine, recombinant clone 4G10® (catalog #05-777) as lot-to-lot variability is essentially eliminated.

      Related Products & Applications

      Alternative Packsize

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

      Alternative Format

      Catalogue 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
      17-123 Anti-Phosphotyrosine Immunoblotting Kit (4G10®, HRP conjugate) Show Pricing & Availability

      Included Positive Control

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

      Product Families

      Categories

      Life Science Research > Antibodies and Assays > Primary Antibodies