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Anti-AREB6/ZEB1 Antibody Picoband™ (monoclonal, 8B12D7)

Figure 1. Western blot analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human U87 whole cell lysates,
Lane 2: rat brain tissue lysates,
Lane 3: mouse brain tissue lysates,
Lane 4: mouse lung tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-AREB6/ZEB1 antigen affinity purified monoclonal antibody (Catalog # M00548-2) at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for AREB6/ZEB1 at approximately 200 kDa. The expected band size for AREB6/ZEB1 is at 124 kDa.
Figure 2. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of human glioblastoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 3. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of human thyroid cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 4. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 5. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 6. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of human laryngeal squamous cell carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 7. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of human lung adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 8. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of human testicular germ cell tumor tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 9. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 10. IHC analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 11. IF analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in a paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®550 Conjugated Avidin (BA1134). Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 12. IF analysis of AREB6/ZEB1 using anti-AREB6/ZEB1 antibody (M00548-2).
AREB6/ZEB1 was detected in an immunocytochemical section of U87 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-AREB6/ZEB1 Antibody (M00548-2) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Mouse IgG (BA1133) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

SPECIFICATION
CITATIONS
PROTOCOLS
BACKGROUND

Product Information
Application Applications Legend: WB=Western Blot IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin-embedded Sections) IHC-F=Immunohistochemistry (Frozen Sections) IF=Immunofluorescence FC=Flow Cytopmetry IC=Immunochemistry ICC=Immunocytochemistry E=ELISA IP=Immunoprecipitation DB=Dot Blot CHIP=Chromatin Immunoprecipitation FA=Fluorescence Assay IEM=Immuno electron microscopy EIA=Enzyme Immunoassay	WB, IHC, IF, ICC
Primary Accession	P37275
Host	Mouse
Isotype	Mouse IgG2b
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-AREB6/ZEB1 Antibody Picoband™ (monoclonal, 8B12D7) . Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Reconstitution	Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Additional Information
Gene ID	6935
Other Names	Zinc finger E-box-binding homeobox 1 {ECO:0000312\|HGNC:HGNC:11642}, NIL-2-A zinc finger protein {ECO:0000303\|Ref.26}, Negative regulator of IL2, Transcription factor 8, TCF-8, ZEB1 (HGNC:11642)
Calculated MW	200 kDa
Application Details	Western blot, 0.25-0.5 µg/ml, Human, Mouse, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human Immunofluorescence, 5 µg/ml, Human
Contents	Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clone Names	Clone: 8B12D7
Immunogen	A synthetic peptide corresponding to a sequence in the middle region of human AREB6/ZEB1.
Purification	Immunogen affinity purified.
Storage	At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Protein Information
Name	ZEB1 (HGNC:11642)
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'). In the absence of TGFB1, acts as a repressor of COL1A2 transcription via binding to the E-box in the upstream enhancer region (By similarity).
Cellular Location	Nucleus
Tissue Location	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

Research Areas

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Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

Background

ZEB1 (Zinc Finger E Box-Binding Homeobox 1), also called TCF8, NIL2A or DELTA-EF1, is a protein that in humans is encoded by the ZEB1 gene. Fluorescence in situ hybridization localized the ZEB1 gene to chromosome 10p11.2. Krafchak et al. (2005) demonstrated a complex (core plus secondary) binding site for TCF8 in the promoter of the COL4A3 gene, mutant in Alport syndrome and which encodes collagen type IV alpha-3. They detected expression of TCF8 in cornea. Nishimura et al. (2006) found that delta-Ef1 was upregulated during differentiation in a mouse smooth muscle cell (SMC) line.

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