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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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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) MSDS

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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