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Products:Cell Biology >> Cell Cycle >> Cell Cycle Inhibitors >> Other
Overview
Product name
Anti-CHFR antibody
See all CHFR products (5) ...
Description
Rabbit polyclonal to CHFR
Specificity
The immunogen for this antibody is identical in all 3 isoforms of CHFR listed in SwissProt ID Q96EP1.
Tested applications
Cross reactivity
Reacts with
Human
Predicted to work with
Mouse
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 450 - 550 of Human CHFR.
(Peptide available as ab24681.)
Positive control
This antibody gave a positive signal in Hela, Jurkat and A431 whole cell lysates
Properties
Form
Liquid
Storage instructions
Store 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 buffer
Preservative: 0.02% Sodium Azide
Constituents: PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
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
Show applications key
Our Abpromise guarantee covers the use of ab4184 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
- 1 Image
WB
Application notes
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 76 kDa (predicted molecular weight: 73 kDa).
This antibody also appears to work in Immunofluoresence - see picture.
We believe this antibody is probably specific for CHFR because in IF with this antibody we see nuclear dots in SAOS2 cells (which have wt CHFR) but not in U2OS cells (that have a mutation in CHFR and hence low levels of it).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
Function
E3 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 specificity
Ubiquitous.
Pathway
Protein modification; protein ubiquitination.
Sequence similarities
Belongs to the CHFR family.
Contains 1 FHA domain.
Contains 1 PBZ-type zinc finger.
Contains 1 RING-type zinc finger.
Developmental stage
Weakly expressed in G1 phase, and highly expressed during S phase.
Domain
The 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
modifications
Poly-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 localization
Nucleus > PML body.
Target information above from: UniProt accessionQ96EP1
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Checkpoint with forkhead and ring finger domains antibody
- Checkpoint with forkhead and RING finger domains protein antibody
- Chfr antibody
see all
Database links
- Clone ID: BC012072 Human
- Entrez Gene: 55743 Human
- Entrez Gene: 231600 Mouse
- IMAGE Id: 4650348 Human
- Omim: 605209 Human
- SwissProt: Q96EP1 Human
- SwissProt: Q810L3 Mouse
- Unigene: 720197 Human
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 BondTM 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.
References for Anti-CHFR antibody (ab4184)
This product has been referenced in:
- Ahel Iet al. Poly(ADP-ribose)-binding zinc finger motifs in DNA repair/checkpoint proteins. Nature 451:81-5 (2008). WB; Human.Read more (PubMed: 18172500) »
See 1 publication for this product
Publishing research using ab4184? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"