1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. heat up to 98 degrees C, below boiling, and then let cool for 10-20 min.
Use at an assay dependent concentration. PubMed: 22123820
Is unsuitable for Flow Cyt.
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.
Expressed predominantly in thymus.
Belongs to the CUT homeobox family. Contains 2 CUT DNA-binding domains. Contains 1 homeobox DNA-binding domain.
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.
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.
There are 2 isoforms produced by alternative splicing.
DNA binding protein SATB1 antibody
DNA-binding protein SATB1 antibody
SATB homeobox 1 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
Western blot - Anti-SATB1 antibody [EPR3951] (ab109122)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg) Lane 2: SATB1 knockout HAP1 whole cell lysate (20 µg) Lane 3: Jurkat whole cell lysate (20 µg) Lane 4: THP1 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109122 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab109122 was shown to specifically react with SATB1 in wild-type HAP1 cells as signal was lost in SATB1 knockout cells. Wild-type and SATB1 knockout samples were subjected to SDS-PAGE. Ab109122 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.
Ab109122 staining SATB1 in Jurkat (Human T cell leukemia T lymphocyte) cells by Immunocytochemistry/Immunofluorescence (ICC/IF). Cells were fixed with 4% paraformaldehyde and permeabilized in 0.1% TritonX-100. Samples were incubated with primary antibody 1:1000 dilution (2.1 μg/ml). An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution (2µg/ml). Alexa Fluor® 594 Anti-alpha Tubulin [DM1A] – Microtubule marker, ab195889 was used as a counterstain antibody (1:200, 2.5 μg/ml). DAPI was used as a counterstain antibody. Confocal image showing nuclear staining on Jurkat cell line.
Western blot - Anti-SATB1 antibody [EPR3951] (ab109122)
All lanes : Anti-SATB1 antibody [EPR3951] (ab109122) at 1/1000 dilution
Lane 1 : Jurkat cell lysates Lane 2 : Fetal thymus lysates Lane 3 : Mouse thymus lysates
Lysates/proteins at 10 µg per lane.
Secondary All lanes : HRP labelled Goat anti-Rabbit at 1/2000 dilution
Ab109122 staining SATB1 in THP-1 (Human monocytic leukemia monocyte) cells by Immunocytochemistry/Immunofluorescence. Cells were fixed with 4% paraformaldehyde and permeabilized in 0.1% TritonX-100. Samples were incubated with primary antibody 1:1000 dilution (2.1 μg/ml). An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution (2µg/ml). Alexa Fluor® 594 Anti-alpha Tubulin [DM1A] – Microtubule marker, ab195889 was used as a counterstain antibody (1:200, 2.5 μg/ml). DAPI was used as a counterstain antibody. Confocal image showing nuclear staining on THP-1 cell line.
ab109122, at 1/100 dilution staining SATB1 in paraffin-embedded Human breast carcinoma, by Immunohistochemistry.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SATB1 antibody [EPR3951] (ab109122)This image is courtesy of an Abreview by Ahmar Aziz
ab109122 staining SATB1 in mouse skin tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue samples were fixed with paraformaldehyde, blocked with 10% goat serum for 30 minutes at 22°C and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/50 in PBS) at 4°C for 12 hours. A FITC-conjugated Goat anti-rabbit polyclonal (1/200) was used as the secondary antibody.
Gong J et al. Hepatic SATB1 induces paracrine activation of hepatic stellate cells and is upregulated by HBx. Sci Rep6:37717 (2016).
Read more (PubMed: 27883059) »
Hao B et al. An anti-silencer- and SATB1-dependent chromatin hub regulates Rag1 and Rag2 gene expression during thymocyte development. J Exp Med212:809-24 (2015).
Read more (PubMed: 25847946) »