RabMAb

Anti-SATB1抗体[EPR3895] (ab92307)

概述

  • 产品名称Anti-SATB1抗体[EPR3895]
    参阅全部 SATB1 一抗
  • 描述
    兔单克隆抗体[EPR3895] to SATB1
  • 经测试应用适用于: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human SATB1 aa 1-100 (N terminal).

  • 阳性对照
    • WB: Jurkat cell lysate and human, rat and mouse thymus tissue lysates. IHC-P: Human colonic carcinoma, thymus and tonsil tissues and rat spleen tissue. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells. IP: Jurkat cell lysate.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Alternative versions available:

    Anti-SATB1 antibody (Alexa Fluor® 488) [EPR3895] (ab204267)

    Anti-SATB1 antibody (Alexa Fluor® 647) [EPR3895] (ab204469)

性能

应用

Our Abpromise guarantee covers the use of ab92307 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000. Predicted molecular weight: 85 kDa.

For unpurified use at 1/1000 - 1/10000.

IP Use at an assay dependent concentration.

For unpurified use at 1/10 - 1/100.

IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/50 - 1/100.

Flow Cyt 1/130.

For unpurified use at 1/1000.

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

ICC/IF 1/250 - 1/500.

For unpurified use at 1/50 - 1/100.

靶标

  • 功能Crucial silencing factor contributing to the initiation of X inactivation mediated by Xist RNA that occurs during embryogenesis and in lymphoma (By similarity). Binds to DNA at special AT-rich sequences, the consensus SATB1-binding sequence (CSBS), at nuclear matrix- or scaffold-associated regions. Thought to recognize the sugar-phosphate structure of double-stranded DNA. Transcriptional repressor controlling nuclear and viral gene expression in a phosphorylated and acetylated status-dependent manner, 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 (e.g. PML at the MHC-I locus) and also by recruiting corepressors (HDACs) or coactivators (HATs) directly to promoters and enhancers. Modulates genes that are essential in the maturation of the immune T-cell CD8SP from thymocytes. Required for the switching of fetal globin species, and beta- and gamma-globin genes regulation during erythroid differentiation. Plays a role in chromatin organization and nuclear architecture during apoptosis. Interacts with the unique region (UR) of cytomegalovirus (CMV). Alu-like motifs and SATB1-binding sites provide a unique chromatin context which seems preferentially targeted by the HIV-1 integration machinery. Moreover, HIV-1 Tat may overcome SATB1-mediated repression of IL2 and IL2RA (interleukin) in T-cells by binding to the same domain than HDAC1. Delineates specific epigenetic modifications at target gene loci, directly upregulating metastasis-associated genes while downregulating tumor-suppressor genes. Reprograms chromatin organization and the transcription profiles of breast tumors to promote growth and metastasis.
  • 组织特异性Expressed predominantly in thymus.
  • 序列相似性Belongs to the CUT homeobox family.
    Contains 2 CUT DNA-binding domains.
    Contains 1 homeobox DNA-binding domain.
  • 翻译后修饰Sumoylated. Sumoylation promotes cleavage by caspases.
    Phosphorylated by PKC. Acetylated by PCAF. Phosphorylated form interacts with HDAC1, but unphosphorylated form interacts with PCAF. DNA binding properties are activated by phosphorylation and inactivated by acetylation. In opposition, gene expression is down-regulated by phosphorylation but up-regulated by acetylation.
    Cleaved at Asp-254 by caspase-3 and caspase-6 during T-cell apoptosis in thymus and during B-cell stimulation. The cleaved forms can not dimerize and lose transcription regulation function because of impaired DNA and chromatin association.
  • 细胞定位Nucleus matrix. Nucleus > PML body. Organized into a cage-like network anchoring loops of heterochromatin and tethering specialized DNA sequences. When sumoylated, localized in promyelocytic leukemia nuclear bodies.
  • Information by UniProt
  • 数据库链接
  • 形式There are 2 isoforms produced by alternative splicing.
  • 别名
    • DNA binding protein SATB1 antibody
    • DNA-binding protein SATB1 antibody
    • SATB homeobox 1 antibody
    • SATB1 antibody
    • SATB1_HUMAN antibody
    • Special AT rich sequence binding protein 1 (binds to nuclear matrix/scaffold associating DNA) antibody
    • Special AT rich sequence binding protein 1 antibody
    • Special AT-rich sequence-binding protein 1 antibody
    see all

Anti-SATB1 antibody [EPR3895] 图像

  • Anti-SATB1 antibody [EPR3895] (ab92307) at 1/1000 dilution (purified) + Human thymus tissue lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 85 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thymus tissue labelling SATB1 with purified ab92307 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling SATB1 with purified ab92307 at 1/500. 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. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

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

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Overlay histogram showing Jurkat cells stained with unpurified ab92307 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92307 (unpurified), 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Unpurified ab92307 immunoprecipitating SATB1 in Jurkat cell lysate.

  • Anti-SATB1 antibody [EPR3895] (ab92307) at 1/1000 dilution (purified) + Rat thymus tissue lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 85 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue labelling SATB1 with purified ab92307 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Unpurified ab92307 staining SATB1 in murine primary thymocytes (isolated from a 21 day old male mouse) by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde, permeabilized using 0.1% Triton X-100 for 5 minutes, blocked with 2% BSA for 1 hour at 25°C and then incubated with unpurified ab92307 at a 1/200 dilution for 3 hours at 25°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • Flow Cytometry analysis of Jurkat cells labelling SATB1 with purified ab92307 at 1/130 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Anti-SATB1 antibody [EPR3895] (ab92307) at 1/10000 dilution (purified) + Mouse thymus tissue lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 85 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human tonisl tissue labelling SATB1 with unpurified ab92307 at a 1/50 dilution.

  • All lanes : Anti-SATB1 antibody [EPR3895] (ab92307) at 1/1000 dilution (unpurified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : Mouse thymus tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 85 kDa

Anti-SATB1 antibody [EPR3895] (ab92307)参考文献

This product has been referenced in:
  • Ma X  et al. Baicalein suppresses metastasis of breast cancer cells by inhibiting EMT via downregulation of SATB1 and Wnt/ß-catenin pathway. Drug Des Devel Ther 10:1419-41 (2016). Human . Read more (PubMed: 27143851) »
  • Peng Z  et al. Co-delivery of doxorubicin and SATB1 shRNA by thermosensitive magnetic cationic liposomes for gastric cancer therapy. PLoS One 9:e92924 (2014). WB ; Human . Read more (PubMed: 24675979) »

See all 8 Publications for this product

Product Wall

Thanks for the update. It at least looks like the antibody has much less affinity for SATB2 than SATB1, assuming equal loading. The other antibody I had suggested was ab109122. (I think I had referred to it as ab92307 at some point, which is what you a...

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The concentration for ab92307, lot GR14706-8, is 0.256 mg/mL.
I hope this is helpful. Please contact me again if you have any further questions.

Of the two rabbit monoclonals, ab92446 (SATB2) and ab92307 (SATB1), ab92446 is more likely to be specific for its target, SATB2, than ab92307 is for SATB1.

Click here (or use the following: http://www.abcam.com/index.html?datasheet=92446). <...

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Thank you for sending the catalogue abID's.

The alignments of the immunogens with the other proteins do suggest that these antibodies may not have the specificity you hope for:

The ab92307 immunogen shares 73% identity (11/15) wit...

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Thank you for contacting Abcam.



The concentration for the current lot of cat# ab92307, is ***********.



Also, a rabbit IgG isotype control is suitable:



http://www.abcam.com/Rabbit-IgG-monoclonal-...

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Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Primary mouse thymocytes)
Specification Primary mouse thymocytes
Fixative Formaldehyde
Permeabilization Yes - 0.1 % TritonX-100 for 5 minutes
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2 · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 14 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293T cells)
Loading amount 20 µg
Specification HEK293T cells
Gel Running Conditions Reduced Denaturing (10 % SDS-PAGE gel)
Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Mar 18 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"