This gene encodes a zinc finger transcription factor. The encoded protein likely plays a role in transcriptional repression of interleukin 2. Mutations in this gene have been associated with posterior polymorphous corneal dystrophy-3 and late-onset Fuchs endothelial corneal dystrophy. Alternatively spliced transcript variants encoding different isoforms have been described.[provided by RefSeq, Mar 2010]
Function:
Acts as a transcriptional repressor. Inhibits interleukin-2 (IL-2) gene expression. Enhances or represses the promoter activity of the ATP1A1 gene depending on the quantity of cDNA and on the cell type. Represses E-cadherin promoter and induces an epithelial-mesenchymal transition (EMT) by recruiting SMARCA4/BRG1. Represses BCL6 transcription in the presence of the corepressor CTBP1. Positively regulates neuronal differentiation. Represses RCOR1 transcription activation during neurogenesis. Represses transcription by binding to the E box (5'-CANNTG-3'). Promotes tumorigenicity by repressing stemness-inhibiting microRNAs.
Subunit:
Interacts (via N-terminus) with SMARCA4/BRG1.
Subcellular Location:
Nucleus.
Tissue Specificity:
Colocalizes with SMARCA4/BRG1 in E-cadherin-negative cells from established lines, and stroma of normal colon as well as in de-differentiated epithelial cells at the invasion front of colorectal carcinomas (at protein level). Expressed in heart and skeletal muscle, but not in liver, spleen, or pancreas.
DISEASE:
Corneal dystrophy, posterior polymorphous, 3 (PPCD3) [MIM:609141]: A subtype of posterior corneal dystrophy, a disease characterized by alterations of Descemet membrane presenting as vesicles, opacities or band-like lesions on slit-lamp examination and specular microscopy. Affected patient typically are asymptomatic. Note=The disease is caused by mutations affecting the gene represented in this entry.
Corneal dystrophy, Fuchs endothelial, 6 (FECD6) [MIM:613270]: A corneal disease caused by loss of endothelium of the central cornea. It is characterized by focal wart-like guttata that arise from Descemet membrane and develop in the central cornea, epithelial blisters, reduced vision and pain. Descemet membrane is thickened by abnormal collagenous deposition. Note=The disease is caused by mutations affecting the gene represented in this entry.
Similarity:
Belongs to the delta-EF1/ZFH-1 C2H2-type zinc-finger family.
Contains 7 C2H2-type zinc fingers.
Contains 1 homeobox DNA-binding domain.
SWISS:
P37275
Gene ID:
6935
Database links:
Entrez Gene: 396029 Chicken
Entrez Gene: 535183 Cow
Entrez Gene: 6935 Human
Entrez Gene: 21417 Mouse
Entrez Gene: 25705 Rat
Omim: 189909 Human
SwissProt: P36197 Chicken
SwissProt: P37275 Human
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Sample:
Kidney (Mouse) Lysate at 40 ug
Primary: Anti- ZEB1/NIL2A (SL4187R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 124 kD
Observed band size: 124 kD
Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti- ZEB1/NIL2A (SL4187R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 124 kD
Observed band size: 124 kD
Sample:
Thymus (Mouse) Lysate at 40 ug
Primary: Anti- ZEB1/NIL2A (SL4187R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 124 kD
Observed band size: 124 kD
Paraformaldehyde-fixed, paraffin embedded (Human breast cancer); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ZEB1) Polyclonal Antibody, Unconjugated (SL4187R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Blank control:Jurkat.
Primary Antibody (green line): Rabbit Anti-ZEB1/NIL2A antibody (SL4187R)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Blank control: H9C2 cells
Isotype Control Antibody: Rabbit IgG-PE ;
Primary Antibody Dilution: 5μl in 100 μL1X PBS containing 0.5% BSA.
Positive control: (mo)H9C2(2% Paraformaldehyde-fixed )
Isotype Control Antibody: Rabbit IgG;
Dilution: 1μg in 100 μl 1 X PBS containing 0.5% BSA
Secondary Antibody: Goat anti-rabbit IgG-FITC;
Dilution: 1:200 in 1 X PBS containing 0.5% BSA
Primary Antibody catalog number: SL4187R; Dilution: 1μg in 100 μl 1X PBS containing 0.5% BSA
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