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Products:Cell Biology >> Cell Cycle >> Cell Division >> Chromatid Cohesion
Overview
Product name
Anti-SMC3 antibody - ChIP Grade
See all SMC3 products (9) ...
Description
Rabbit polyclonal to SMC3 - ChIP Grade
Tested applications
ICC, IP, WB, ChIP, IHC-Pmore details
Cross reactivity
Reacts with
Mouse, Human, Xenopus laevis
Predicted to work with
Cow
Immunogen
Synthetic peptide (Human) conjugated to KLH - which represents a portion within the last 100 amino acids of the human Structural Maintenance of Chromosomes-3 (GenBank PID 4885399), conjugated toKLH.
Positive control
HeLa nuclear or whole cell extract
Properties
Form
Liquid
Storage instructions
Store at +4°C.
Storage buffer
Preservative: 0.09% Sodium Azide
Constituents: 8mM PBS, 60mM Citrate, 150mM Tris, pH 7-8
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
Antibodies were affinity purified using the peptide immobilized on solid support. Antibody concentration was determined by extinction coefficient: absorbance at 280 nmof 1.4 equals 1.0 mg of IgG.
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 >> BRCA1
Cell Biology >> Cell Cycle >> Cell Division >> Chromatid Cohesion
Applications
Show applications key
Our Abpromise guarantee covers the use of ab9263 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC: 1/200 - 1/800.
IP: Use a concentration of 2 - 6 µg/ml.
WB: 1/1000 - 1/10000.Detects a band of approximately 140 kDa (predicted molecular weight: 141.5 kDa).
ChIP: Use at an assay dependent dilution. (PubMed: 20966046)
IHC-P: Use at an assay dependent dilution.
Target
Function
Central component of cohesin, a complex required for chromosome cohesion during the cell cycle. The cohesin complex may form a large proteinaceous ring within which sister chromatids can be trapped. At anaphase, the complex is cleaved and dissociates from chromatin, allowing sister chromatids to segregate. Cohesion is coupled to DNA replication and is involved in DNA repair. The cohesin complex plays also an important role in spindle pole assembly during mitosis and in chromosomes movement.
Involvement in disease
Defects in SMC3 are the cause of Cornelia de Lange syndrome type 3 (CDLS3) [MIM:610759]. CDLS is a dominantly inherited multisystem developmental disorder characterized by growth and cognitive retardation, abnormalities of the upper limbs, gastroesophageal dysfunction, cardiac, ophthalmologic and genitourinary anomalies, hirsutism, and characteristic facial features. CDSL3 is a mild form with absence of major structural anomalies typically associated with CDLS. The phenotype in some instances approaches that of apparently non-syndromic mental retardation.
Sequence similarities
Belongs to the SMC family. SMC3 subfamily.
Domain
The flexible hinge domain, which separates the large intramolecular coiled coil regions, allows the heterotypic interaction with the corresponding domain of SMC1A or SMC1B, forming a V-shaped heterodimer. The two heads of the heterodimer are then connected by different ends of the cleavable RAD21 protein, forming a ring structure.
Post-translational
modifications
Phosphorylated upon DNA damage, probably by ATM or ATR.
Acetylation at Lys-105 and Lys-106 by ESCO1 is important for genome stability and S phase sister chromatid cohesion. Regulated by DSCC1, it is required for processive DNA synthesis, coupling sister chromatid cohesion establishment during S phase to DNA replication.
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, the RAD21 subunit of the cohesin complex is cleaved, leading to the dissociation of the complex from chromosomes, allowing chromosome separation.
Target information above from: UniProt accessionQ9UQE7
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Basement membrane associated chondroitin proteoglycan antibody
- Chromosome associated polypeptide antibody
- BAM antibody
see all
Database links
- Entrez Gene: 281729 Cow
- Entrez Gene: 9126 Human
- Entrez Gene: 13006 Mouse
- Entrez Gene: 399092 Xenopus laevis
- Omim: 606062 Human
- SwissProt: Q9UQE7 Human
- SwissProt: Q9CW03 Mouse
- SwissProt: O93309 Xenopus laevis
see all
Anti-SMC3 antibody - ChIP Grade images:
Immunocytochemistry - SMC3 antibody (ab9263)
SMC3 signals (green) in a human spermatocyte at the zygotene-pachytene transition stage. Wider signals correspond to synapsed areas, whereas narrower signals show non-synapsed regions.
This image was kindly supplied as part of the review submitted by Maria Oliver-Bonet.
Immunocytochemistry - SMC3 antibody (ab9263)
Interphase HeLa cells stained with ab9263 (1/500). The antibody gave predominantly nuclear staining in all interphase cells investigated. ab9263 staining is shown in green. The cells were counterstained with DAPI (red).
Kirk McManus, University of British Columbia
Western blot - SMC3 antibody (ab9263)
All lanes : Anti-SMC3 antibody - ChIP Grade (ab9263) at 1 µg/ml
Lane 1 : Placenta (Human) Tissue Lysate - adult normal tissue (ab29745)
Lane 2 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like 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
Predicted band size : 141.5 kDa
Observed band size : 145 kDa (why is the actual band size different from the predicted?)
Immunoprecipitation - SMC3 antibody (ab9263)
ab9263 Immunoprecipitate in human HeLa cytosolic and nuclear extracts. 500µg of cell lysate incubated with primary antibody (1/100) and matrix (Protein A) in 20mM HEPES ph 7.8, 100mM KAc for 4 hours at 4°C. For Western blotting ab9263 was used at a dilution at 1/1000.
This image is a courtesy of Anonymous Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-SMC3 antibody(ab9263)
ab9263 (4µg/ml) staining SMC3 in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
References for Anti-SMC3 antibody - ChIP Grade (ab9263)
This product has been referenced in:
- Ishiguro Ket al. A new meiosis-specific cohesin complex implicated in the cohesin code for homologous pairing. EMBO Rep 12:267-75 (2011). WB; Mouse.Read more (PubMed: 21274006) »
- Nativio Ret al. Disruption of genomic neighbourhood at the imprinted IGF2-H19 locus in Beckwith-Wiedemann syndrome and Silver-Russell syndrome. Hum Mol Genet 20:1363-74 (2011). ChIP; Human.Read more (PubMed: 21282187) »
See all 8 publications for this product
Publishing research using ab9263? 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"