Anti-CHFR antibody (ab4184)

Overview

• Product nameAnti-CHFR antibodySee all CHFR primary antibodies ...
• Description
  Rabbit polyclonal to CHFR
• SpecificityThe immunogen for this antibody is identical in all 3 isoforms of CHFR listed in SwissProt ID Q96EP1.
• Tested applicationsWB more details
• Species reactivity
  Reacts with: Human
  Predicted to work with: Mouse
• Immunogen

  Synthetic peptide conjugated to KLH derived from within residues 450 - 550 of Human CHFR.

  Read Abcam's proprietary immunogen policy

  (Peptide available as ab24681.)

• Positive controlThis antibody gave a positive signal in Hela, Jurkat and A431 whole cell lysates

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C (add glycerol to a final volume of 50% for extra stability). Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: PBS, pH 7.4
• Concentration information loading...
• PurityImmunogen affinity purified
• Clonality Polyclonal
• IsotypeIgG
• Research Areas
  • Cancer
  • Cell cycle
  • Cell cycle inhibitors
  • Other

  • Epigenetics and Nuclear Signaling
  • Cell cycle
  • Cell Cycle Inhibitors
  • Other

  • Cell Biology
  • Cell Cycle
  • Cell Cycle Inhibitors
  • Other

Applications

Target

• FunctionE3 ubiquitin-protein ligase that functions in the antephase checkpoint by actively delaying passage into mitosis in response to microtubule poisons. Acts in early prophase before chromosome condensation, when the centrosome move apart from each other along the periphery of the nucleus. Probably involved in signaling the presence of mitotic stress caused by microtubule poisons by mediating the 'Lys-48'-linked ubiquitination of target proteins, leading to their degradation by the proteasome. Promotes the ubiquitination and subsequent degradation of AURKA and PLK1. Probably acts as a tumor suppressor, possibly by mediating the polyubiquitination of HDAC1, leading to its degradation. May also promote the formation of 'Lys-63'-linked polyubiquitin chains and functions with the specific ubiquitin-conjugating UBC13-MMS2 (UBE2N-UBE2V2) heterodimer. Substrates that are polyubiquitinated at 'Lys-63' are usually not targeted for degradation, but are rather involved in signaling cellular stress.
• Tissue specificityUbiquitous.
• PathwayProtein modification; protein ubiquitination.
• Sequence similaritiesBelongs to the CHFR family.
  Contains 1 FHA domain.
  Contains 1 PBZ-type zinc finger.
  Contains 1 RING-type zinc finger.
• Developmental stageWeakly expressed in G1 phase, and highly expressed during S phase.
• DomainThe PBZ-type zinc finger (also named CYR) mediates non-covalent poly(ADP-ribose)-binding. Poly(ADP-ribose)-binding is dependent on the presence of zinc and is required for its function in antephase checkpoint.
  The FHA domain plays a key role in the anti-proliferative properties of the protein and is involved in initiating a cell cycle arrest at G2/M. The FHA domain may be required to interact with phosphorylated proteins.
• Post-translational
  modificationsPoly-ADP-ribosylated. In addition to binding non covalently poly(ADP-ribose) via its PBZ-type zinc finger, the protein is also covalently poly-ADP-ribosylated by PARP1.
  Autoubiquitinated; may regulate its cellular level.
  Phosphorylated upon DNA damage, probably by ATM or ATR (By similarity). Phosphorylated by PKB. Phosphorylation may affect its E3 ligase activity.
• Cellular localizationNucleus > PML body.

Target information above from: UniProt accession Q96EP1 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 55743 Human
  • Entrez Gene: 55743 Human
  • Entrez Gene: 231600 Mouse
  • Omim: 605209 Human
  • SwissProt: Q96EP1 Human
  • SwissProt: Q96EP1 Human
  • SwissProt: Q810L3 Mouse
  • Unigene: 720197 Human

  • Unigene: 30264 Mouse

  see all

• Alternative names
  Checkpoint with forkhead and ring finger domains antibodyCheckpoint with forkhead and RING finger domains protein antibodyChfr antibody

  CHFR_HUMAN antibodyE3 ubiquitin protein ligase CHFR antibodyE3 ubiquitin-protein ligase CHFR antibodyEC 6.3.2. antibodyFLJ10796 antibodyFLJ33629 antibodyRING finger protein 196 antibodyRNF 116 antibodyRNF 196 antibodyRNF116 antibodyRNF196 antibodyUbiquitin ligase protein antibody

  see all

Anti-CHFR antibody images

• 

  Immunofluorescence - Anti-CHFR antibody (ab4184)

  Immunofluoresence using ab4184 on SAOS2 cells with a FITC conjugated secondary antibody.

  The foci seen are confined to the nuclei of the cells present in the picture.

• 

  Western blot - CHFR antibody (ab4184)
  All lanes : Anti-CHFR antibody (ab4184) at 1 µg/ml
  
  Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
  Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
  Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size : 73 kDa
  Observed band size : 76 kDa (why is the actual band size different from the predicted?)
  Additional bands at : 34 kDa. We are unsure as to the identity of these extra bands.
• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHFR antibody (ab4184)
  IHC image of ab4184 staining in Human Breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab4184, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  
  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.