Anti-SMC1A antibody (ab21583)
Key features and details
- Rabbit polyclonal to SMC1A
- Suitable for: ICC/IF, WB, IP, IHC-P
- Reacts with: Human
- 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: ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Chicken, Xenopus laevis
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Immunogen
Synthetic peptide corresponding to Human SMC1A aa 1200 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab23863) -
Positive control
- HeLa and Jurkat whole cell lysate
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab21583 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 ICC/IF Use a concentration of 1 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 150 kDa (predicted molecular weight: 143 kDa). IP Use at an assay dependent concentration. IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. 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: 8243 Human
- Entrez Gene: 24061 Mouse
- Entrez Gene: 63996 Rat
- Omim: 300040 Human
- SwissProt: Q14683 Human
- SwissProt: Q9CU62 Mouse
- SwissProt: Q9Z1M9 Rat
- SwissProt: O93308 Xenopus laevis
see all -
Alternative names
- Chromosome segregation protein SmcB antibody
- DXS423E antibody
- KIAA0178 antibody
see all
Images
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Western blot - Anti-SMC1A antibody (ab21583)All lanes : Anti-SMC1A antibody (ab21583) at 1 µg/ml
Lane 1 : 20ug HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899) at 20 µg
Lane 3 : 20ug HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate withHuman SMC1A peptide (ab23863) at 1 µg/ml
Lane 4 :Jurkat whole cell lysate (ab7899) at 20 µg with Human SMC1A peptide (ab23863) at 1 µg/ml
Secondary
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) (ab28446) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 143 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?ab21583 detects a band of 150 kDa, corresponding to the size of SMC1A. This band is competed away by the addition of the immunizing peptide, showing that it is a specific interaction.
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Immunoprecipitation - Anti-SMC1A antibody (ab21583)SMC1A was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to SMC1A and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 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 ab21583.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 150kDa: SMC1A; Non specific - 41 and 42kDa: We are unsure as to the identity of this extra band. -
Immunocytochemistry/ Immunofluorescence - Anti-SMC1A antibody (ab21583)ICC/IF image of ab21583 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21583, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Panel A shows localisation of ab21583 to the nuclei, Panel B has the Alexa Fluor® 488 channel removed for comparison.
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Immunocytochemistry/ Immunofluorescence - Anti-SMC1A antibody (ab21583)This image is courtesy of an Abreview submitted by Dr Kirk McManusab21583 staining SMC1A in assynchonous HeLa cells (green). Cells were paraformaldehyde-fixed (4% - 10min) and counterstained with DAPI (red). Secondary antibody: Goat anti-Rabbit conjugated to Cy3 ®. Please refer to Abreview for further details.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMC1A antibody (ab21583)IHC image of SMC1A staining in human skin FFPE section, performed on a Leica BondTM 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 ab21583, 1µg/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.
Protocols
Datasheets and documents
References (12)
ab21583 has been referenced in 12 publications.
- Dufey E et al. Genotoxic stress triggers the activation of IRE1a-dependent RNA decay to modulate the DNA damage response. Nat Commun 11:2401 (2020). WB ; Mouse . PubMed: 32409639
- Prati B et al. Three Prime Repair Exonuclease 1 (TREX1) expression correlates with cervical cancer cells growth in vitro and disease progression in vivo. Sci Rep 9:351 (2019). PubMed: 30674977
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
- Bellelli R et al. Pole Instability Drives Replication Stress, Abnormal Development, and Tumorigenesis. Mol Cell 70:707-721.e7 (2018). PubMed: 29754823
- Zhang Y et al. Phosphorylation of SMC1A promotes hepatocellular carcinoma cell proliferation and migration. Int J Biol Sci 14:1081-1089 (2018). PubMed: 29988990
Images
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Western blot - Anti-SMC1A antibody (ab21583)All lanes : Anti-SMC1A antibody (ab21583) at 1 µg/ml
Lane 1 : 20ug HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899) at 20 µg
Lane 3 : 20ug HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate withHuman SMC1A peptide (ab23863) at 1 µg/ml
Lane 4 :Jurkat whole cell lysate (ab7899) at 20 µg with Human SMC1A peptide (ab23863) at 1 µg/ml
Secondary
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) (ab28446) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 143 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?ab21583 detects a band of 150 kDa, corresponding to the size of SMC1A. This band is competed away by the addition of the immunizing peptide, showing that it is a specific interaction.
-
Immunoprecipitation - Anti-SMC1A antibody (ab21583)
SMC1A was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to SMC1A and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 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 ab21583.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 150kDa: SMC1A; Non specific - 41 and 42kDa: We are unsure as to the identity of this extra band. -
Immunocytochemistry/ Immunofluorescence - Anti-SMC1A antibody (ab21583)
ICC/IF image of ab21583 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21583, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Panel A shows localisation of ab21583 to the nuclei, Panel B has the Alexa Fluor® 488 channel removed for comparison.
-
Immunocytochemistry/ Immunofluorescence - Anti-SMC1A antibody (ab21583) This image is courtesy of an Abreview submitted by Dr Kirk McManus
ab21583 staining SMC1A in assynchonous HeLa cells (green). Cells were paraformaldehyde-fixed (4% - 10min) and counterstained with DAPI (red). Secondary antibody: Goat anti-Rabbit conjugated to Cy3 ®. Please refer to Abreview for further details.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMC1A antibody (ab21583)
IHC image of SMC1A staining in human skin FFPE section, performed on a Leica BondTM 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 ab21583, 1µg/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.
