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Products:Cell Biology >> Cell Cycle >> Cell Division >> Chromatid Cohesion
Overview
Product name
Anti-Rad21 antibody - ChIP Grade
See all Rad21 products (5) ...
Description
Rabbit polyclonal to Rad21 - ChIP Grade
Tested applications
ChIP, ICC/IF, WB, IPmore details
Cross reactivity
Reacts with
Mouse, Human, Xenopus laevis, Indian Muntjac
Immunogen
Synthetic peptide (Human) conjugated to KLH - which represented a portion of human Rad21 encoded within exon 14 (LocusLink ID 5885).
Positive control
HeLa cell lysate
Properties
Form
Liquid
Storage instructions
Store at +4°C.
Storage buffer
Preservative: 0.1% Sodium Azide
Constituents: 8mM PBS, 60mM Citrate, 150mM Tris, pH 7-8
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Cancer >> Cell cycle >> Cell division
Epigenetics and Nuclear Signaling >> Cell cycle >> Chromosome Structure >> Chromatid Cohesion
Epigenetics and Nuclear Signaling >> Chromosome Structure >> Cohesins
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Damage & Repair >> DNA Damage Response >> Other
Cell Biology >> Cell Cycle >> Cell Division >> Chromatid Cohesion
Applications
Show applications key
Our Abpromise guarantee covers the use of ab992 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ChIP: Use at an assay dependent dilution.
ICC/IF: Use at an assay dependent concentration. (Used at a dilution of 1/200 for 30 min incubation (see Abreview for further information).)
WB: 1/500 - 1/5000.Predicted molecular weight: 72 kDa.
IP: Use a concentration of 1 - 4 µg/ml.
Target
Function
Cleavable component of the cohesin complex, involved in chromosome cohesion during cell cycle, in DNA repair, and in apoptosis. The cohesin complex is required for the cohesion of sister chromatids after DNA replication. The cohesin complex apparently forms a large proteinaceous ring within which sister chromatids can be trapped. At metaphase-anaphase transition, this protein is cleaved by separase/ESPL1 and dissociates from chromatin, allowing sister chromatids to segregate. The cohesin complex may also play a role in spindle pole assembly during mitosis. Also plays a role in apoptosis, via its cleavage by caspase-3/CASP3 or caspase-7/CASP7 during early steps of apoptosis: the C-terminal 64 kDa cleavage product may act as a nuclear signal to initiate cytoplasmic events involved in the apoptotic pathway.
Sequence similarities
Belongs to the rad21 family.
Domain
The C-terminal part associates with the head of SMC1A, while the N-terminal part binds to the head of SMC3.
Post-translational
modifications
Cleaved by separase/ESPL1 at the onset of anaphase. Cleaved by caspase-3 and caspase-7 at the beginning of apoptosis. The cleavage by ESPL1 and caspase-3 take place at different sites.
Phosphorylated; becomes hyperphosphorylated in M phase of cell cycle. The large dissociation of cohesin from chromosome arms during prophase may be partly due to its phosphorylation by PLK.
Cellular localization
Nucleus. Chromosome. Chromosome > centromere. Associates with chromatin. Before prophase it is scattered along chromosome arms. During prophase, most of cohesin complexes dissociate from chromatin probably because of phosphorylation by PLK, except at centromeres, where cohesin complexes remain. At anaphase, it is cleaved by separase/ESPL1, leading to the dissociation of the complex from chromosomes, allowing chromosome separation. Once cleaved by caspase-3, the C-terminal 64 kDa cleavage product translocates to the cytoplasm, where it may trigger apoptosis.
Target information above from: UniProt accessionO60216
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Double strand break repair protein rad21 homolog antibody
- Double-strand-break repair protein rad21 homolog antibody
- FLJ25655 antibody
see all
Database links
- Entrez Gene: 5885 Human
- Entrez Gene: 19357 Mouse
- Entrez Gene: 399129 Xenopus laevis
- Omim: 606462 Human
- SwissProt: O60216 Human
- SwissProt: Q61550 Mouse
- SwissProt: O93310 Xenopus laevis
- Unigene: 81848 Human
see all
Anti-Rad21 antibody - ChIP Grade images:
Western blot - Rad21 antibody - ChIP Grade (ab992)
Anti-Rad21 antibody - ChIP Grade (ab992) at 1/1000 dilution
Predicted band size : 72 kDa
Observed band size : 75 kDa (why is the actual band size different from the predicted?)
Additional bands at : 130 kDa (possible cross reactivity).
Western blot - Rad21 antibody (ab992)
Anti-Rad21 antibody - ChIP Grade (ab992) at 1/2000 dilution + HeLa whole cell extract. Extraction was achieved using RIPA buffer and 1% SDS. 25 µg of total protein was loaded
Secondary
Goat anti rabbit IgG (HRP) at 1/1500 dilution
Predicted band size : 72 kDa
A rad21 knock down negative control was employed.
This image is courtesy of an Abreview submitted on 9 September 2005. We do not have any further information relating to this image.
Immunocytochemistry/ Immunofluorescence - Rad21 antibody (ab992)
ab992 at a 1/200 dilution staining Asynchronous and paraformaldehyde-fixed (4%) HeLa cells by immunocytochemistry. The antibody was incubated with the cells 30 minutes and then detected using a Cy3 conjugated Goat Anti-Mouse IgG (H+L) antibody.
This image is courtesy of an Abreview by Kirk McManus submitted on 27 February 2006.
Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
All lanes : Anti-Rad21 antibody - ChIP Grade (ab992) at 2 µg/ml
Lane 1 : HeLa at 50 µg
Lane 2 : HeLa at 15 µg
Lane 3 : HeLa at 5 µg
Lane 4 : 293T at 50 µg
Lane 5 : NIH3T3 at 50 µg
Predicted band size : 72 kDa
Exposure time : 30 seconds
References for Anti-Rad21 antibody - ChIP Grade (ab992)
This product has been referenced in:
- Xiao Tet al. Specific Sites in the C Terminus of CTCF Interact with the SA2 Subunit of the Cohesin Complex and Are Required for Cohesin-Dependent Insulation Activity. Mol Cell Biol 31:2174-83 (2011). WB; Human.Read more (PubMed: 21444719) »
- Chatterjee Set al. Dynamic changes in binding of immunoglobulin heavy chain 3' regulatory region to protein factors during class switching. J Biol Chem 286:29303-12 (2011). ChIP; Mouse.Read more (PubMed: 21685395) »
See all 18 publications for this product
Publishing research using ab992? 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"