Our Abpromise guarantee covers the use of ab34735 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Notes
WB
WB: Use a concentration of 1 µg/ml. Detects a band of approximay 85 kDa (predicted molecular weight: 83 kDa).Can be blocked with Mouse SATB2 peptide (ab34906).
EMSA
EMSA: Use at an assay dependent concentration. PubMed: 22123820
IHC-FoFr
IHC-FoFr: Use at an assay dependent dilution.
ICC/IF
ICC/IF: Use at an assay dependent concentration. PubMed: 19232401
Target
FunctionBinds 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 specificityHigh 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 diseaseNote=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 similaritiesBelongs to the CUT homeobox family. Contains 2 CUT DNA-binding domains. Contains 1 homeobox DNA-binding domain.
Post-translational modificationsSumoylated by PIAS1. Sumoylation promotes nuclear localization, but represses transcription factor activity.
Cellular localizationNucleus matrix.
Alternative names
DNA binding protein SATB2 antibody
DNA-binding protein SATB2 antibody
FLJ21474 antibody
see all
Anti-SATB2 antibody images
Western blot - SATB2 antibody (ab34735)
Anti-SATB2 antibody (ab34735) at 1 µg/ml + Brain (Mouse) Tissue Lysate - normal tissue, 0 days old (ab7188) at 10 µg
Predicted band size : 83 kDa Observed band size : 85 kDa (why is the actual band size different from the predicted?) Additional bands at : 30 kDa. We are unsure as to the identity of these extra bands.
Electrophoretic Mobility Shift Assay - Anti-SATB2 antibody (ab34735)Image from Asanoma K et al, J Biol Chem. 2012 Jan 13;287(3):2257-68. Epub 2011 Nov 28, Fig 8. DOI 10.1074/jbc.M111.287128 January 13, 2012 The Journal of Biological Chemistry, 287, 2257-2268.
ab34735 used in EMSA. EMSA demonstrating binding of SATB1 and SATB2 to an oligonucleotide containing a predicted SATB-binding element located 125 bp upstream of the Eomes transcription start site (-125). Labeled SATB probes were incubated with Rcho-1 TS cell nuclear lysates as indicated above the lanes. Lane 1, without any competitor oligonucleotide; lane 2, with unlabeled wild-type competitor; lane 3, with unlabeled mutant competitor; lane 4, with SATB1 antibody; lane 5, with SATB2 antibody (ab34735); lane 6, with a control antibody. B and C, Rcho-1 TS cells stably transfected with pcDNA3, pcDNA3-HA-Satb1, or pcDNA3-HA-Satb2 and maintained in stem culture conditions were used for ChIP assays. Antibodies to the HA tag (Anti-HA) or acetylated histone H3K9 (Anti-AcH3) were used in the analyses. IgG was used as a control for nonspecific immunoprecipitation. qPCR analyses were performed on immunoprecipitates using primers amplifying the segment (-172 to -63) including the SATB-binding element at -125.
Immunocytochemistry/ Immunofluorescence - Anti-SATB2 antibody (ab34735)Image courtesy of an anonymous Abreview.
ab34735 staining SATB2 in human osteosarcoma SAOS-2 cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde, permeabilized using PBS/ 0.25% Triton, blocked with 1% BSA for 1 hour at room temperature and then incubated with ab34735 at a 1/500 dilution for 1 hour. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/250 dilution.
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