Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-SATB2 antibody [EPNCIR130A] - BSA and Azide free (ab212177)

Overview

  • Product name
    Anti-SATB2 antibody [EPNCIR130A] - BSA and Azide free
    See all SATB2 primary antibodies
  • Description
    Rabbit monoclonal [EPNCIR130A] to SATB2 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IF, Flow Cyt, IHC-Frmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human SATB2. The exact sequence is proprietary.

  • Positive control
    • WB: HT-1080, SW1353, MCF7 and Saos-2 cell lysates. Rat and mouse brain and human fetal brain tissue lysates. IHC-P: Human cerebral cortex tissue. ICC/IF: SH-SY5Y cells.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of John Niederhuber. View antibodies from NCI Center for Cancer Research Collaboration.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab212177 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 Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

WB Use at an assay dependent concentration. Predicted molecular weight: 81 kDa.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Binds to DNA, at nuclear matrix- or scaffold-associated regions. Thought to recognize the sugar-phosphate structure of double-stranded DNA. Transcription factor controlling nuclear gene expression, by binding to matrix attachment regions (MARs) of DNA and inducing a local chromatin-loop remodeling. Acts as a docking site for several chromatin remodeling enzymes and also by recruiting corepressors (HDACs) or coactivators (HATs) directly to promoters and enhancers. Required for the initiation of the upper-layer neurons (UL1) specific genetic program and for the inactivation of deep-layer neurons (DL) and UL2 specific genes, probably by modulating BCL11B expression. Repressor of Ctip2 and regulatory determinant of corticocortical connections in the developing cerebral cortex. May play an important role in palate formation. Acts as a molecular node in a transcriptional network regulating skeletal development and osteoblast differentiation.
    • Tissue specificity
      High expression in adult brain, moderate expression in fetal brain, and weak expression in adult liver, kidney, and spinal cord and in select brain regions, including amygdala, corpus callosum, caudate nucleus, and hippocampus.
    • Involvement in disease
      Note=Chromosomal aberrations involving SATB2 are found in isolated cleft palate. Translocation t(2;7); translocation t(2;11).
      Defects in SATB2 are a cause of cleft palate isolated (CPI) [MIM:119540]. A congenital fissure of the soft and/or hard palate, due to faulty fusion. Isolated cleft palate is not associated with cleft lips. Some patients may manifest other craniofacial dysmorphic features, mental retardation, and osteoporosis.
      Note=A chromosomal aberration involving SATB2 is found in a patient with classical features of Toriello-Carey syndrome. Translocation t(2;14)(q33;q22).
    • Sequence similarities
      Belongs to the CUT homeobox family.
      Contains 2 CUT DNA-binding domains.
      Contains 1 homeobox DNA-binding domain.
    • Post-translational
      modifications
      Sumoylated by PIAS1. Sumoylation promotes nuclear localization, but represses transcription factor activity.
    • Cellular localization
      Nucleus matrix.
    • Information by UniProt
    • Database links
    • Alternative names
      • DNA binding protein SATB2 antibody
      • DNA-binding protein SATB2 antibody
      • FLJ21474 antibody
      • FLJ32076 antibody
      • GLSS antibody
      • KIAA1034 antibody
      • MGC119474 antibody
      • MGC119477 antibody
      • SATB family member 2 antibody
      • SATB homeobox 2 antibody
      • SATB2 antibody
      • SATB2_HUMAN antibody
      • Special AT rich sequence binding protein 2 antibody
      • Special AT-rich sequence-binding protein 2 antibody
      see all

    Images

    • All lanes : Anti-SATB2 antibody [EPNCIR130A] (ab92446) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : SATB2 knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 81 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab92446 observed at 83 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab92446 was shown to specifically react with SATB2 in wild-type HAP1 cells as signal was lost in SATB2 knockout cells. Wild-type and SATB2 knockout samples were subjected to SDS-PAGE. Ab92446 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92446).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue labelling SATB2 with purified ab92446 at 1/150. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92446).

    • Flow Cytometry analysis of SH-SY5Y cells (human neuroblastoma cell line from bone marrow)  labeling SATB2 with purified ab92446 at 1/150 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde  and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92446).

    • Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y (human neuroblastoma cell line from bone marrow) cells labelling SATB2 (green) with purified ab92446 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/100) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92446).

    • Unpurified ab92446 staining SATB2 in E18 Mouse brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.3% Triton-X 100 and blocked with 3% BSA for 30 minutes at 25°C. The sample was incubated with primary antibody (1/500 in TBS with 0.1% Triton-X 100 + 3% Goat serum) at 4°C for 12 hours. An Alexa Fluor® 546-conjugated Goat anti-rabbit polyclonal (1/1000) was used as the secondary antibody.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92446).

    • Unpurified ab92446 staining SATB2 in mouse brain tissue by Immunohistochemistry (Frozen sections). Tissue was fixed with paraformaldehyde and permeabilized using 0.3% Triton-X-100. Samples were then blocked with 5% serum for 1 hour 30 minutes at 20°C followed by incubation with the primary antibody at a 1/200 dilution for 36 hours. A biotin conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/200 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92446).

    References

    ab212177 has not yet been referenced specifically in any publications.

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