Anti-SMC1A antibody (ab9262)
Key features and details
- Rabbit polyclonal to SMC1A
- Suitable for: IHC-P, ICC, IP, WB
- Reacts with: Mouse, Human, Xenopus laevis
- Isotype: IgG
Overview
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Product name
Anti-SMC1A antibody
See all SMC1A primary antibodies -
Description
Rabbit polyclonal to SMC1A -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Human, Xenopus laevis
Predicted to work with: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Pig, Rhesus monkey, Gorilla, Orangutan, Platypus
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Immunogen
Synthetic peptide within Human SMC1A aa 1133-1233 conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
Database link: Q14683
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris -
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 280nm of 1.4 equals 1.0 mg of IgG. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab9262 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P | 1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
| ICC | 1/200 - 1/800. | |
| IP | Use at 1-4 µg/mg of lysate. | |
| WB | 1/1000 - 1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 143 kDa). |
Target
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Function
Involved 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 disease
Defects 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 similarities
Belongs to the SMC family. SMC1 subfamily. -
Domain
The 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. -
Post-translational
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 localization
Nucleus. 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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Database links
- Entrez Gene: 282370 Cow
- Entrez Gene: 8243 Human
- Entrez Gene: 24061 Mouse
- Entrez Gene: 63996 Rat
- Omim: 300040 Human
- SwissProt: O97593 Cow
- SwissProt: Q14683 Human
- SwissProt: Q9CU62 Mouse
see all -
Alternative names
- Chromosome segregation protein SmcB antibody
- DXS423E antibody
- KIAA0178 antibody
see all
Images
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Immunocytochemistry - Anti-SMC1A antibody (ab9262)This image is courtesy of Kirk McManus, University of British ColumbiaInterphase HeLa cells stained with ab9262 (1/500). The antibody gave nuclear staining in all interphase nuclei investigated. However, the antibody failed to recognize any chromatin-associated epitopes in prophase or metaphase cells suggesting that the epitopes may be masked during mitosis. ab9262 staining is shown in green. The cells were counterstained with DAPI (red). 100x magnification.
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Immunoprecipitation - Anti-SMC1A antibody (ab9262)Immunoprecipitation analysis of HeLa whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) prepared using NETN lysis buffer.
Lanes 1 & 2: ab9262 was used for IP at 6 µg per reaction.
Lane 3: IP using rabbit anti-SMC1A antibody (ab140493).
Lane 4: Control IgG.For western blotting immunoprecipitated SMC1, ab9262 was used at 1 µg/ml.
Detection: Chemiluminescence. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMC1A antibody (ab9262)Ab9262 staining human normal colon tissue. Staining is localised to nuclear compartment.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
References (30)
ab9262 has been referenced in 30 publications.
- Pickel C et al. Oxygen-dependent bond formation with FIH regulates the activity of the client protein OTUB1. Redox Biol 26:101265 (2019). PubMed: 31299612
- Korsholm LM et al. Double-strand breaks in ribosomal RNA genes activate a distinct signaling and chromatin response to facilitate nucleolar restructuring and repair. Nucleic Acids Res 47:8019-8035 (2019). PubMed: 31184714
- Leimbacher PA et al. MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis. Mol Cell 74:571-583.e8 (2019). PubMed: 30898438
- Li J et al. SMC1 promotes proliferation and inhibits apoptosis through the NF-?B signaling pathway in colorectal cancer. Oncol Rep N/A:N/A (2019). PubMed: 31524239
- Mehta K & Laimins L Human Papillomaviruses Preferentially Recruit DNA Repair Factors to Viral Genomes for Rapid Repair and Amplification. MBio 9:N/A (2018). PubMed: 29440569
