Key features and details
- Rabbit polyclonal to SMC3
- Suitable for: IP, WB, IHC-P, ICC/IF
- Reacts with: Mouse, Human, Xenopus laevis
- Isotype: IgG
Product nameAnti-SMC3 antibody
See all SMC3 primary antibodies
DescriptionRabbit polyclonal to SMC3
Tested applicationsSuitable for: IP, WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Human, Xenopus laevis
Predicted to work with: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Turkey, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan, Platypus
Synthetic peptide corresponding to Human SMC3 conjugated to keyhole limpet haemocyanin. Which represents a portion within the last 100 amino acids GenBank PID 4885399.
- WB: Human placenta tissue lysate. K562 and Jurkat whole cell lysate. ICC/IF: HeLa cells. Human spermatocyte. IP: SMC3 IP in nuclear extracts from HeLa cells. IHC-P: Human colon and testicular seminoma tissue.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.09% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
Concentration information loading...
PurityImmunogen affinity purified
Purification notesAntibodies 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.
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.
|IP||Use at 2-10 µg/mg of lysate.|
|WB||1/1000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 141.5 kDa).|
|IHC-P||1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||1/200 - 1/800.|
FunctionCentral 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 diseaseDefects 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 similaritiesBelongs to the SMC family. SMC3 subfamily.
DomainThe 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.
modificationsPhosphorylated 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 localizationNucleus. 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.
- Information by UniProt
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SMC3 is phosphorylated during prophase I.
(Part of panel B) Nuclear spreads of spermatocytes were labeled with anti-pS1083, anti-SMC3 (ab9263) and anti-SYCP3 antibodies. Arrowheads indicate the XY bivalent. Bars, 10 µm.
All lanes : Anti-SMC3 antibody (ab9263) at 1 µg/ml
Lane 1 : Human placenta tissue lysate - total protein (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.
All lanes : 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?
Interphase HeLa (Human epithelial cell line from cervix adenocarcinoma) 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).
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.
ab9263 Immunoprecipitate in HeLa (Human epithelial cell line from cervix adenocarcinoma) cytosolic and nuclear extracts.
500 µg of cell lysate incubated with primary antibody (1/100) and matrix (Protein A) in 20mM HEPES ph 7.8, 100 mM KAc for 4 hours at 4°C.
For Western blotting ab9263 was used at a dilution at 1/1000.
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 3-in-1 AR buffer citrate pH 6.0. Slides were blocked with peroxidase in 3% H2O2 in methanol for 10 minutes. They were then blocked with a protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with an amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with hematoxylin 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testicular seminoma tissue labeling SMC3 with ab9263 at 1/1000 ( 1µg/ml). Detection: DAB.
ab9263 has been referenced in 52 publications.
- Leylek TR et al. Reduced Expression of Genes Regulating Cohesion Induces Chromosome Instability that May Promote Cancer and Impact Patient Outcomes. Sci Rep 10:592 (2020). PubMed: 31953484
- Zielinska AP et al. Meiotic Kinetochores Fragment into Multiple Lobes upon Cohesin Loss in Aging Eggs. Curr Biol 29:3749-3765.e7 (2019). PubMed: 31679939
- Wang T et al. Smc3 is required for mouse embryonic and adult hematopoiesis. Exp Hematol 70:70-84.e6 (2019). PubMed: 30553776
- Furlan C et al. Miniaturised interaction proteomics on a microfluidic platform with ultra-low input requirements. Nat Commun 10:1525 (2019). PubMed: 30948724
- Full F et al. Centrosomal protein TRIM43 restricts herpesvirus infection by regulating nuclear lamina integrity. Nat Microbiol 4:164-176 (2019). PubMed: 30420784