Key features and details
- Rabbit polyclonal to SMC1A (phospho S966)
- Suitable for: IHC-P, IP, WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Product nameAnti-SMC1A (phospho S966) antibody
See all SMC1A primary antibodies
DescriptionRabbit polyclonal to SMC1A (phospho S966)
Tested applicationsSuitable for: IHC-P, IP, WB, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Rhesus monkey, Orangutan
Synthetic peptide corresponding to SMC1A conjugated to keyhole limpet haemocyanin.
- HeLa cells treated with 10 Gy IR for 1hr. IHC-P: FFPE human spleen tissue sections.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
Concentration information loading...
PurityImmunogen affinity purified
Purification notesAntibodies that were not phospho-specific were removed by solid phase absorption. Antibodies specific for SMC1A pSer957 were affinity purified using the phosphopeptide immobilized onsolid support.
Our Abpromise guarantee covers the use of ab1276 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|IP||Use at 2-6 µg/mg of lysate.|
|WB||1/500 - 1/5000. Detects a band of approximately 160 kDa (predicted molecular weight: 143 kDa).|
|ICC/IF||Use at an assay dependent concentration.|
FunctionInvolved in chromosome cohesion during cell cycle and in DNA repair. Central component of cohesin complex. 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 anaphase, the complex is cleaved and dissociates from chromatin, allowing sister chromatids to segregate. The cohesin complex may also play a role in spindle pole assembly during mitosis. Involved in DNA repair via its interaction with BRCA1 and its related phosphorylation by ATM, or via its phosphorylation by ATR. Works as a downstream effector both in the ATM/NBS1 branch and in the ATR/MSH2 branch of S-phase checkpoint.
Involvement in diseaseDefects in SMC1A are the cause of Cornelia de Lange syndrome type 2 (CDLS2) [MIM:300590]; also known as Cornelia de Lange syndrome X-linked. CDLS is a clinically heterogeneous developmental disorder associated with malformations affecting multiple systems. CDLS is characterized by facial dysmorphisms, abnormal hands and feet, growth delay, cognitive retardation and various other malformations including gastroesophageal dysfunction and cardiac, ophthalmologic and genitourinary anomalies.
Sequence similaritiesBelongs to the SMC family. SMC1 subfamily.
DomainThe flexible hinge domain, which separates the large intramolecular coiled coil regions, allows the heterotypic interaction with the corresponding domain of SMC3, 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 by ATM upon ionizing radiation in a NBS1-dependent manner. Phosphorylated by ATR upon DNA methylation in a MSH2/MSH6-dependent manner. Phosphorylation of Ser-957 and Ser-966 activates it and is required for S-phase checkpoint activation.
Cellular localizationNucleus. Chromosome. Chromosome > centromere > kinetochore. 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. In germ cells, cohesin complex dissociates from chromatin at prophase I, and may be replaced by a meiosis-specific cohesin complex. The phosphorylated form on Ser-957 and Ser-966 associates with chromatin during G1/S/G2 phases but not during M phase, suggesting that phosphorylation does not regulate cohesin function. Integral component of the functional centromere-kinetochore complex at the kinetochore region during mitosis.
- Information by UniProt
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ab1276 at a 1:1000 dilution staining phospho SMC1A (Ser 966) in 100µg of treated (10 Gy IR, 1h) HeLa nuclear extract by Western Blot (ECL). ab1276 at a 1:1000 dilution staining phospho SMC1A (Ser 966) in 100µg of treated (10 Gy IR, 1h) HeLa nuclear extract by Western Blot (ECL).
Immunocytochemistry/Immunofluorescence analysis of neocarzinostatin treated asynchronous HeLa cells labelling SMC1A (phospho S966) with ab1276 at 1/5000 (0.2µg/ml). A DyLight® 594-conjugated anti-rabbit IgG (1/100) was used as the secondary antibody.
IHC image of SMC1A (phospho S966) staining in human spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1276, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
SKN cells cultured on coverslips were fixed in 4% paraformaldehyde and then stained with Rabbit polyclonal to SMC1A (phospho S966), ab1276 (green) at a working dilution of 1/200. The DNA stained with DAPI is shown in red. (100x magnification).
ab1276 has been referenced in 16 publications.
- Leimbacher PA et al. MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis. Mol Cell 74:571-583.e8 (2019). PubMed: 30898438
- Zhang Y et al. Phosphorylation of SMC1A promotes hepatocellular carcinoma cell proliferation and migration. Int J Biol Sci 14:1081-1089 (2018). PubMed: 29988990
- Yang T et al. Quercetin-3-O-ß-D-glucoside decreases the bioavailability of cyclosporin A through regulation of drug metabolizing enzymes, transporters and nuclear receptors in rats. Mol Med Rep 18:2599-2612 (2018). PubMed: 30015887
- Jiang Y et al. ATM function and its relationship with ATM gene mutations in chronic lymphocytic leukemia with the recurrent deletion (11q22.3-23.2). Blood Cancer J 6:e465 (2016). PubMed: 27588518
- Buscemi G et al. Bimodal regulation of p21(waf1) protein as function of DNA damage levels. Cell Cycle 13:2901-12 (2014). WB . PubMed: 25486478