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Products:Epigenetics and Nuclear Signaling >> Histones >> Variants
Overview
Product name
Anti-CENPA (di methyl K9) antibody
See all CENPA products (10) ...
Description
Rabbit polyclonal to CENPA (di methyl K9)
Specificity
This antibody is blocked by the modified peptide (ab13488) but not the unmodified peptide.
Tested applications
Cross reactivity
Reacts with
Cow, Human
Immunogen
Synthetic peptide derived from within residues 1 - 100 of Human CENPA, di methylated at K9.
(Peptide available as ab13488.)
Positive control
This antibody gave a positive signal in HeLa nuclear and calf thymus histone preparations.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
-
Western blot
(enlarge)
Applications
Show applications key
Our Abpromise guarantee covers the use of ab4488 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 17 kDa (predicted molecular weight: 16 kDa). Can be blocked with CENPA peptide - di methyl K9 (ab13488).
We have had customer reports that this antibody does not appear to be suitable for ICC/IF in either HeLa, SK-N-SH or 10T1/2 cells. ab4488 should only stain at the centromeres, however in colocalization studies with ACA (anti-centromeric antigen), ab4488 was distributed throughout the nucleus and did not colocalised with ACA.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
Function
Histone H3-like variant which exclusively replaces conventional H3 in the nucleosome core of centromeric chromatin at the inner plate of the kinetochore. Required for recruitment and assembly of kinetochore proteins, mitotic progression and chromosome segregation. May serve as an epigenetic mark that propagates centromere identity through replication and cell division.
Sequence similarities
Belongs to the histone H3 family.
Domain
The CATD (CENPA targeting domain) region is responsible for the more compact structure of nucleosomes containing CENPA and is necessary and sufficient to mediate the localization into centromeres.
Post-translational
modifications
Ubiquitinated (Probable). Interaction with herpes virus HSV-1 ICP0 protein, leads to its degradation by the proteasome pathway.
Phosphorylation of Ser-7 by Aurora-A/STK6 and Aurora-B/STK12 during prophase is required for localization of Aurora-A/STK6 and Aurora-B/STK12 at inner centromere and is essential for kinetochore function. Initial phosphorylation during prophase is mediated by Aurora-A/STK6 and is maintained by Aurora-B/STK12.
Cellular localization
Nucleus. Chromosome > centromere > kinetochore. Localizes exclusively in the kinetochore domain of centromeres. Occupies a compact domain at the inner kinetochore plate stretching across 2 thirds of the length of the constriction but encompassing only one third of the constriction width and height.
Target information above from: UniProt accessionP49450
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- CENP A antibody
- CENP-A antibody
- cenpa antibody
see all
Database links
- Entrez Gene: 1058 Human
- Omim: 117139 Human
- SwissProt: P49450 Human
- Unigene: 1594 Human
Anti-CENPA (di methyl K9) antibody images:
Western blot
All lanes : Anti-CENPA (di methyl K9) antibody (ab4488) at 1/500 dilution
Lane 1 : HeLa Cell Lysate at 20 µg
Lane 2 : HeLa nuclear lysate at 20 µg
Lane 3 : Calf thymus histone prep at 1 µg
Lane 4 : HeLa Cell Lysate at 20 µg with CENPA peptide - di methyl K9 (ab13488) at 1 µg/ml
Lane 5 : HeLa nuclear lysate at 20 µg with CENPA peptide - di methyl K9 (ab13488) at 1 µg/ml
Lane 6 : Calf thymus histone prep at 1 µg with CENPA peptide - di methyl K9 (ab13488) at 1 µg/ml
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size : 16 kDa
Observed band size : 18 kDa (why is the actual band size different from the predicted?)
Additional bands at : 50 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 30 seconds
Rabbit polyclonal to CENPA (di methyl K9) at 1/500.
Lane 1 HeLa cell lysate (20ug)
Lane 2 HeLa nuclear lysate (20ug)
Lane 3 Calf thymus histone prep (1ug)
Lane 4 HeLa cell lysate (20ug) - blocking peptide (ab13488)
Lane 5 HeLa nuclear lysate (20ug) - blocking peptide (ab13488)
Lane 6 Calf thymus histone prep (1ug) - blocking peptide (ab13488)
Secondary antibody - Goat polyclonal to Rabbit IgG (ab6721) at 1/5000.
References for Anti-CENPA (di methyl K9) antibody (ab4488)
ab4488 has not yet been referenced specifically in any publications.
Publishing research using ab4488? 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"