Loading...
You have changed your country from
to
. Please be aware that this will change the currency in the purchasing process.
Products:Cell Biology >> Cell Cycle >> Cell Division >> Chromatid Cohesion
Overview
Product name
Anti-Rad21 antibody
See all Rad21 products (5) ...
Description
Rabbit polyclonal to Rad21
Tested applications
WB, ICC/IF, IPmore details
Cross reactivity
Reacts with
Mouse, Human
Predicted to work with
Rat, Chicken, Cow
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human Rad21.
(Peptide available as ab42520.)
Positive control
This antibody gave a positive signal in the following lysates: HeLa Whole Cell, HEK293 Whole Cell, A431 Whole Cell, Mouse Brain Tissue, Mouse Testis Tissue
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
Research areas
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 ab42478 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 100 kDa (predicted molecular weight: 75 kDa).(Although the predicted band size is 75kDa based on Swiss-prot data, a band of 120kDa has been previously observed. Mol Cell Biol. 2002 Dec;22(23):8267-77 PMID: 12417729. The band seen at 100kDa is also consistent with the banding pattern observed for other commercially available antibodies to Rad21.)
ICC/IF: Use a concentration of 1 µg/ml
IP: Use at an assay dependent concentration.
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
- hHR21 antibody
- HR21 antibody
see all
Database links
- Entrez Gene: 420286 Chicken
- Entrez Gene: 540966 Cow
- Entrez Gene: 5885 Human
- Entrez Gene: 19357 Mouse
- Omim: 606462 Human
- SwissProt: Q3SWX9 Cow
- SwissProt: O60216 Human
- SwissProt: Q61550 Mouse
see all
Anti-Rad21 antibody images:
Immunocytochemistry/ Immunofluorescence - Rad21 antibody (ab42478)
ab42478 (1/200) staining Rad21 in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100 and counterstained with DAPI in order to highlight the nucleus (red). for further experimental details please see Abreview.
Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
Western blot - Rad21 antibody (ab42478)
All lanes : Anti-Rad21 antibody (ab42478) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 75 kDa
Observed band size : 100 kDa (why is the actual band size different from the predicted?)
Additional bands at : 55 kDa (possible cleavage fragment).
Although the predicted band size is 75kDa based on Swiss-prot data, a band of 120kDa has been previously observed. Mol Cell Biol. 2002 Dec;22(23):8267-77 PMID: 12417729. The band seen at 100kDa is also consistent with the banding pattern observed for other commercially available antibodies to Rad21.
Immunocytochemistry/ Immunofluorescence - Rad21 antibody (ab42478)
ICC/IF image of ab42478 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab42478, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Western blot - Rad21 antibody (ab42478)
All lanes : Anti-Rad21 antibody (ab42478) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate
Lane 2 : Testis (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 75 kDa
Observed band size : 120 kDa (why is the actual band size different from the predicted?)
Additional bands at : 55 kDa,85 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 4 minutes
Although the predicted band size is 75kDa based on Swiss-prot data, a band of 120kDa has been previously observed. Mol Cell Biol. 2002 Dec;22(23):8267-77 PMID: 12417729.
Immunoprecipitation - Anti-Rad21 antibody (ab42478)
Rad21 was immunoprecipitated using 0.5mg Hela whole cell extract, 5ug of Rabbit polyclonal to Rad21and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extractdiluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab42478. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) .. Band: 100kDa: Rad21; Non specific - 50kDa: We are unsure as to the identity of this extra band. Although the predicted band size is 75kDa based on Swiss-prot data, a band of 120kDa has been previously observed. Mol Cell Biol. 2002 Dec;22(23):8267-77 PMID: 12417729. The band seen at 100kDa is also consistent with the banding pattern observed for other commercially available antibodies to Rad21.
References for Anti-Rad21 antibody (ab42478)
ab42478 has not yet been referenced specifically in any publications.
Publishing research using ab42478? 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"