Overview

  • Product name
    Anti-SMC3 antibody - ChIP Grade
    See all SMC3 primary antibodies
  • Description
    Rabbit polyclonal to SMC3 - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, WB, ChIP, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts 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
  • Immunogen

    Synthetic peptide corresponding to Human SMC3 conjugated to keyhole limpet haemocyanin. Which represents a portion within the last 100 amino acids GenBank PID 4885399.

  • Positive control
    • 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.

Properties

Applications

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.

Application Abreviews Notes
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).
ChIP Use at an assay dependent concentration. PubMed: 20966046
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.

Target

  • Function
    Central 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 disease
    Defects 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 similarities
    Belongs to the SMC family. SMC3 subfamily.
  • Domain
    The 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.
  • Post-translational
    modifications
    Phosphorylated 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 localization
    Nucleus. 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
  • Database links
  • Alternative names
    • BAM antibody
    • Bamacan antibody
    • Basement membrane associated chondroitin proteoglycan antibody
    • Basement membrane-associated chondroitin proteoglycan antibody
    • BMH antibody
    • CDLS3 antibody
    • chondroitin sulfate proteoglycan 6 (bamacan) antibody
    • Chondroitin sulfate proteoglycan 6 antibody
    • Chromosome associated polypeptide antibody
    • Chromosome-associated polypeptide antibody
    • CSPG 6 antibody
    • CSPG6 antibody
    • hCAP antibody
    • Human chromosome associated polypeptide antibody
    • im:7142991 antibody
    • SMC 3 antibody
    • SMC protein 3 antibody
    • SMC-3 antibody
    • smc3 antibody
    • SMC3_HUMAN antibody
    • SMC3L1 antibody
    • Structural maintenance of chromosome 3 antibody
    • Structural maintenance of chromosomes 3 antibody
    • Structural maintenance of chromosomes protein 3 antibody
    • u:fb22e01 antibody
    • wu:fc30d07 antibody
    see all

Images

  • 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 - ChIP Grade (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.

    Secondary
    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.

    See Abreview

  • 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.

References

This product has been referenced in:
  • Tsai PF  et al. A Muscle-Specific Enhancer RNA Mediates Cohesin Recruitment and Regulates Transcription In trans. Mol Cell 71:129-141.e8 (2018). Read more (PubMed: 29979962) »
  • Casey AE  et al. Differential cohesin loading marks paired and unpaired regions of platypus sex chromosomes at prophase I. Sci Rep 7:4217 (2017). Read more (PubMed: 28652620) »
See all 39 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Scorpion Cell (spermatocyte at in pachytene)
Permeabilization
No
Specification
spermatocyte at in pachytene
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Mr. Bruno Almeida

Verified customer

Submitted Apr 27 2017

Answer


I have performed an alignment of the sequence provided in the word document with the sequence used for ab9263. At the site recognized by ab9263, the human and Schistosoma mansoni (blood fluke) sequences are too divergent to expect ab9263 to bind.

Please let me know however if you would like to test ab74582 in Schistosoma mansoni.

Read More

Answer



Unfortunately, at the site recognized by ab9263, the human and Schistosoma mansoni sequences are too divergent to expect ab9263 to bind.

However, Anti-SMC3 antibody's (ab74582) immunogen shows a 90% sequence homology with the above sequence and therefore could work in Schistosoma mansoni.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela)
Loading amount
50 µg
Specification
Hela
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 01 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa cytosolic and nuclear extracts)
Total protein in input
500 µg
Specification
HeLa cytosolic and nuclear extracts
Immuno-precipitation step
Protein A

Abcam user community

Verified customer

Submitted Nov 03 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cytosolic and nuclear extracts)
Loading amount
50 µg
Specification
HeLa cytosolic and nuclear extracts
Gel Running Conditions
Reduced Denaturing (8% gel)
Blocking step
Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Nov 02 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Dec 07 2005

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Xenopus laevis Purified protein (Chromatin isolated from Xenopus egg extract)
Specification
Chromatin isolated from Xenopus egg extract
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5

Dr. Alan Score

Verified customer

Submitted Nov 28 2005

Answer

Ab3600 - Rabbit polyclonal to Argonaute 4 - has not been discontinued; it is currently in stock in our office in the UK. If you have any more questions, please contact me again.

Read More

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