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ABS1513-I Anti-phospho-Ubiquitin (Ser65)

Detect ubiquitin Ser65 phosphorylation using this rabbit polyclonal Anti-phospho-Ubiquitin (Ser65), Cat. No. ABS1513-I, validated for use in Immunocytochemistry and Western Blotting.

Anti-phospho-Ubiquitin (Ser65) MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Signaling Gene Symbol: UBB, UBC UniProt Number: P0CG47, P0CG478
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ABS1513-I
100 µL  
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      Overview

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      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      HWB, ICCRbAffinity PurifiedPolyclonal Antibody
      Description
      Catalogue NumberABS1513-I
      ReplacesABS1513
      DescriptionAnti-phospho-Ubiquitin (Ser65)
      Alternate Names
      • Ubiqutin, Ser65 phosphorylated
      • Ub, Ser65 phosphorylated
      Background InformationUbiqutin (Ub) is initially produced as a 229-a.a. Polyubiquitin-B (UniProt P0CG47) precursor protein encoded by the UBB gene (Gene ID 7314) or a 685-a.a. Polyubiquitin-C precursor protein (UniProt P0CG48) encoded by the UBC gene (Gene ID 7316) in human. The C-termini of Polyubiquitin-B (Cys229) and Polyubiquitin-C (Val685) are post-translationally removed, followed by additional posttranslational cleavages to yield multiple copies of idential 76-amino acid ubiquitin (also known as high mobility group nonhistone protein HMG-20) molecules (three copies per Polyubiquitin-B and nine copies per Polyubiquitin-C). Covalent modifications of target proteins via different Ub Lys residues are well documented events during cellular DNA repair, ER-associated degradation (ERAD), transcription activation, lysosomal and proteasomal degradation. PINK1 (PTEN induced putative kinase 1) is a Ser/Thr kinase that specifically accumulates in depolarized mitochondria, where it acts as a positive regulator of parkin (Park2) E3 ubiquitin (Ub) ligase activity by phosphorylating Ub at Ser65 as well as the corresponding Ser residue in parkin N-terminal Ub-like (UBL) domain. Phosphorylated Ub interacts with phosphorylated parkin at its GINGO0 domain in an allosteric manner, inducing a conformation change and exposing parkin RING2 domain catalytic cysteine for full ligase activity. NMR based conformation study reveals that Ser65-phosphorylated Ub exists in two conformations in solution. The major conformation displays altered surface properties, while the second phosphoUb conformation exhibits a retracted C-terminal tail by two residues into the Ub core. Ub Ser65 phosphorylation has little effect toward E1-mediated E2 charging and mainly affects the discharging of E2 enzymes to form polyUb chains. In addition, the majority of deubiquitinases (DUBs), including USP8, USP15 and USP30, are shown to exhibit impaired activity against Lys63-linked poly-phosphoUb.
      References
      Product Information
      FormatAffinity Purified
      PresentationPurified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
      Quality LevelMQ100
      Applications
      ApplicationDetect ubiquitin Ser65 phosphorylation using this rabbit polyclonal Anti-phospho-Ubiquitin (Ser65), Cat. No. ABS1513-I, validated for use in Immunocytochemistry and Western Blotting.
      Key Applications
      • Western Blotting
      • Immunocytochemistry
      Application NotesImmunocytochemistry Analysis: A 1:500 dilution from a representative lot detected CCCP (Cat. No. 215911) treatment-induced ubiquitin Ser65 phosphorylation in PINK1-expressing HeLa cells.

      Western Blotting Analysis: A 1:5,000 dilution from a representative lot detected GST-PINK1-catalyzed Ser65 phosphorylation of recombinant Ubiquitin.

      Immunocytochemistry Analysis: A 1:200 dilution from a representative lot detected phospho-Ubiquitin (Ser65) in PINK1 expressing HeLa cells without and with CCCP treatment. (Courtesy of Dr. Noriyuki Matsuda).
      Biological Information
      ImmunogenKLH-conjugated linear peptide corresponding to the ubiquitin target region sequence containing phosphorylated Ser65.
      ConcentrationPlease refer to lot specific datasheet.
      HostRabbit
      SpecificityThis polyclonal antibody detects only Ser65-phosphorylated ubiquitin (Ub) and ubiquitinated proteins (including oligoUb and polyUb) containing Ser65-phosphorylated ubiquitin. No reactivity toward Ub or ubiquitinated proteins in the absence of Ub Ser65 phosphorylation.
      Species Reactivity
      • Human
      Species Reactivity NoteHuman. Expected to react with a broad-range of species, including Bovine, Guinea Pig, Hamster, Horse, Mouse, Non-Human Primate, Rat, Sheep, based on 100% sequence homology.
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • UBB
      • UBC
      Modifications
      • Phosphorylation
      Purification MethodAffinity purified.
      UniProt Number
      Molecular Weight~17/8 and ~45-250 kDa observed. 8.565/17.112/25.66/34.21 kDa (mono-/di/tri-/tetra-ubiquitin) calculated. Bands ~45-250 kDa represent ubiquitinated proteins containing ser65-phosphorylated Ub. Uncharacterized bands may be observed in some lysate(s).
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by Western Blotting in PINK1-expressing HeLa cells.

      Western Blotting Analysis: A 1:1,000 dilution of this antibody detected cellular ubiquitin Ser65 phosphorylation induction in 10 µg of lysate from PINK1-expressing HeLa cells upon CCCP (Cat. No. 215911) treatment.
      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.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      ABS1513-I 04054839077494

      Documentation

      Anti-phospho-Ubiquitin (Ser65) SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-phospho-Ubiquitin (Ser65) Certificates of Analysis

      TitleLot Number
      Anti-phospho-Ubiquitin (Ser65) - 3156000 3156000
      Anti-phospho-Ubiquitin (Ser65) - 3175967 3175967
      Anti-phospho-Ubiquitin (Ser65) - 3221963 3221963
      Anti-phospho-Ubiquitin (Ser65) - 3291392 3291392
      Anti-phospho-Ubiquitin (Ser65) - 3536100 3536100
      Anti-phospho-Ubiquitin (Ser65) - 3587100 3587100
      Anti-phospho-Ubiquitin (Ser65) - 3764707 3764707
      Anti-phospho-Ubiquitin (Ser65) - 3845000 3845000
      Anti-phospho-Ubiquitin (Ser65) - 3898671 3898671
      Anti-phospho-Ubiquitin (Ser65) - 3937801 3937801
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