Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-SATB2 antibody [EPNCIR130A] (ab92446)

Overview

  • Product name
    Anti-SATB2 antibody [EPNCIR130A]
    See all SATB2 primary antibodies
  • Description
    Rabbit monoclonal [EPNCIR130A] to SATB2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, 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.
    Database link: Q9UPW6

  • 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; Mouse brain tissue. ICC/IF: SH-SY5Y cells. Flow Cyt: SH-SY5Y cells.
  • General notes

    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.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92446 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
WB 1/1000 - 1/5000. Predicted molecular weight: 81 kDa.
IHC-P 1/150 - 1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/100.
Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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

    • 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.

      See Abreview

    • 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.

    • 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.

      See Abreview

    • 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).

    • Anti-SATB2 antibody [EPNCIR130A] (ab92446) at 1/1000 dilution (purified) + SW1353 cell lysate at 20 µg

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 81 kDa
      Observed band size: 83 kDa
      why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • 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. 

    • 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.

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

      Lane 1 : Saos-2 (human osteosarcoma cell line) cell lysate
      Lane 2 : Human fetal brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 81 kDa
      Observed band size: 83 kDa why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

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

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 81 kDa
      Observed band size: 83 kDa why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

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

      Lane 1 : HT1080 (human fibrosarcoma cell line) cell lysate
      Lane 2 : SW1353 cell lysate
      Lane 3 : MCF7 (human breast adenocarcinoma cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 81 kDa

    References

    This product has been referenced in:
    • D'Gama AM  et al. Somatic Mutations Activating the mTOR Pathway in Dorsal Telencephalic Progenitors Cause a Continuum of Cortical Dysplasias. Cell Rep 21:3754-3766 (2017). Read more (PubMed: 29281825) »
    • Feng J  et al. BMP signaling orchestrates a transcriptional network to control the fate of mesenchymal stem cells in mice. Development 144:2560-2569 (2017). Read more (PubMed: 28576771) »
    See all 9 Publications for this product

    Customer reviews and Q&As

    1-6 of 6 Q&A

    Question
    Answer

    Thank you for your call yesterday.
    I now heard back form the source of this antibody and I am very happy to confirm that we can provide a range of amino acids where the immunogen in a part of.
    Unfortunately the exact sequence is proprietary and was not communicated to me.
    The immunogen peptide maps between amino acids 240 and 275 of human SATB2 (UniProt: Q9UPW6).
    I hope this information will be helpful to the interpretation of your data.

    Read More

    Answer

    Thank you for contacting us. The concentration of ab51502 is 0.1 mg/ml and should be used at a dilution of 1/100 for both IP and WB. Regarding ab92445, this is a tissue culture supernatant antibody so the concentration is not determined. Generally concentrations are only determined for purified antibodies as they are not relevant for other types that may contains proteins other than the protein of interest. This product should be used at a dilution of 1/10-1/100 for IP and 1/1000-1/10000 for WB. We do not test our antibodies side by side and cannot say which works "better" for a particular application but we do guarantee them both to work in IP and WB.



    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Answer

    Thank you for contacting us. I am very sorry to hear that you are having problems with this antibody. Could you please let me know a few more experimental details (for our records): 1) What sample type was used and what is the species of the samples? 2) What antibody dilution was tried? 3) Which blocking reagent was used? 4) Which secondary antibody/detection system was employed? Also, please let me know your order number. If the item was purchased within the last 6 months, I'd be happy to discuss a replacement with you. Please let me know the requested information. I look forward to hear back from you and assist in resolving this case for you.

    Read More

    Answer

    Thank you for your email. I am very glad to hear that you obtained the desired results. As for another SATB2 antibody that is suitable for IP/WB, we have also ab92446: https://www.abcam.com/SATB2-antibody-EPNCIR130A-ab92446.html It was tested in IP and WB and several other applications, as well as in human, mouse and rat samples. The antibody you are currently using is also suitable for IP: ab51502 Other antibodies tested in WB and human samples are: ab69995, ab69998 and ab119118. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. 

    Read More

    Answer

    Thank you for your reply. Unfortunately, the specific immunogen sequence is proprietary and I am unable to disclose this information.  All other available information regarding this product is available on the online datasheet. I hope this information is helpful and I apologize for any inconvenience caused.   Please do not hesitate to contact us if you have any additional questions.

    Read More

    Answer

    Thank you for contacting Abcam regarding ab92446. I have checked our catalog and unfortunately we do not have a blocking peptide available for this antibody. I apologize for any inconvenience this may cause.  Please do not hesitate to contact us if you have any additional questions.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
    For licensing inquiries, please contact partnerships@abcam.com

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