Overview

  • Product name

  • Description

    Rabbit polyclonal to SMC1A
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Xenopus laevis
  • Immunogen

    Synthetic peptide corresponding to Human SMC1A aa 1200 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab23863)

  • Positive control

    • HeLa and Jurkat whole cell lysate

Properties

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

  • 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
    modifications

    Phosphorylated 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
  • Database links

  • Alternative names

    • Chromosome segregation protein SmcB antibody
    • DXS423E antibody
    • KIAA0178 antibody
    • MGC138332 antibody
    • Sb1.8 antibody
    • Segregation of mitotic chromosomes 1 antibody
    • SMC protein 1A antibody
    • SMC-1-alpha antibody
    • SMC-1A antibody
    • SMC1 (structural maintenance of chromosomes 1 yeast) like 1 antibody
    • SMC1 antibody
    • SMC1 structural maintenance of chromosomes 1 like 1 antibody
    • SMC1A antibody
    • SMC1A_HUMAN antibody
    • SMC1alpha antibody
    • SMC1L1 antibody
    • SMCB antibody
    • Structural maintenance of chromosomes 1A antibody
    • Structural maintenance of chromosomes protein 1A antibody
    see all

Images

  • 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 with Human 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.

  • 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.

  • 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.

  • 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.

    See Abreview

  • 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.

References

This product has been referenced in:

  • 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). Read more (PubMed: 30674977) »
  • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). Read more (PubMed: 30377371) »
See all 11 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse stem cell)
Specification
mouse stem cell
Fixative
Formaldehyde
Permeabilization
Yes - triton
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 18°C

Abcam user community

Verified customer

Submitted Sep 02 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 20 2007

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