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Product Info Summary
| SKU: | A00548-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-AREB6/ZEB1 Antibody Picoband®
SKU/Catalog Number
A00548-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-AREB6/ZEB1 Antibody Picoband® catalog # A00548-2. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store 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 freeze-thaw cycles.
Cite This Product
Anti-AREB6/ZEB1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00548-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of human ZEB1, which shares 94.9% and 100% amino acid (aa) sequence identity with rat ZEB1.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00548-2 is reactive to ZEB1 in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
200 kDa
Calculated molecular weight
9072 MW
Background of ZEB1
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.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00548-2 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human 293T whole cell, human PC-3 whole cell, human HeLa whole cell
IHC: human glioma tissue, human melanoma tissue
ICC/IF: U20S cell
FCM: U2OS cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of ZEB1 using anti-ZEB1 antibody (A00548-2).
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 293T whole cell lysates,
Lane 2: human PC-3 whole cell lysates,
Lane 3: human HeLa whole cell 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 rabbit anti-ZEB1 antigen affinity purified polyclonal antibody (Catalog # A00548-2) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ZEB1 at approximately 200 kDa. The expected band size for ZEB1 is at 124 kDa.
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Figure 2. IHC analysis of ZEB1 using anti-ZEB1 antibody (A00548-2).
ZEB1 was detected in paraffin-embedded section of human glioma tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ZEB1 Antibody (A00548-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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Figure 3. IHC analysis of ZEB1 using anti-ZEB1 antibody (A00548-2).
ZEB1 was detected in paraffin-embedded section of human melanoma tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ZEB1 Antibody (A00548-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IF analysis of ZEB1 using anti-ZEB1 antibody (A00548-2)
ZEB1 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/mL rabbit anti-ZEB1 Antibody (A00548-2) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 5. Flow Cytometry analysis of K562 cells using anti-ZEB1 antibody (A00548-2).
Overlay histogram showing K562 cells stained with A00548-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ZEB1 Antibody (A00548-2,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For ZEB1 (Source: Uniprot.org, NCBI)
Gene Name
ZEB1
Full Name
Zinc finger E-box-binding homeobox 1
Weight
9072 MW
Superfamily
delta-EF1/ZFH-1 C2H2-type zinc-finger family
Alternative Names
Zinc finger E-box-binding homeobox 1; NIL-2-A zinc finger protein; Negative regulator of IL2; Transcription factor 8; TCF-8; ZEB1; AREB6; TCF8 ZEB1 AREB6, BZP, DELTAEF1, FECD6, NIL2A, PPCD3, TCF8, ZFHEP, ZFHX1A zinc finger E-box binding homeobox 1 zinc finger E-box-binding homeobox 1|delta-crystallin enhancer binding factor 1|negative regulator of IL2|posterior polymorphous corneal dystrophy 3|transcription factor 8 (represses interleukin 2 expression)|zinc finger homeodomain enhancer-binding protein
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on ZEB1, check out the ZEB1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for ZEB1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-AREB6/ZEB1 Antibody Picoband® (A00548-2)
Hello CJ!
A00548-2 has been cited in 3 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
All-trans retinoic acid reverses malignant biological behavior of hepatocarcinoma cells by regulating miR-200 family members
MiR-181a contributes gefitinib resistance in non-small cell lung cancer cells by targeting GAS7
Wang Q,Shang J,Zhang Y,Zhou Y,Tang L.MiR-451a restrains the growth and metastatic phenotypes of papillary thyroid carcinoma cells via inhibiting ZEB1.Biomed Pharmacother.2020 Jul;127:109901.doi:10.1016/j. biopha.2020.109901.Epub 2020 Apr 23.PMID:32335297.
Species: Human,Mouse
A00548-2 usage in article: APP:IF, SAMPLE:SW579 CELL AND B-CPAP CELL, DILUTION:1:500
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Customer Reviews
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Customer Q&As
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5 Customer Q&As for Anti-AREB6/ZEB1 Antibody Picoband®
Question
I am looking for using your anti-AREB6/ZEB1 antibody for positive regulation of neuron differentiation studies. Has this antibody been tested with western blotting on human hek293 whole cell lysates? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2017-12-26
Answer
We appreciate your inquiry. This A00548-2 anti-AREB6/ZEB1 antibody is validated on human hek293 whole cell lysates. It is guaranteed to work for Flow Cytometry, IF, IHC-P, ICC, WB in human. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2017-12-26
Question
We have been able to see staining in human myometrium. What should we do? Is anti-AREB6/ZEB1 antibody supposed to stain myometrium positively?
Verified Customer
Verified customer
Asked: 2017-09-21
Answer
From what I have seen in literature myometrium does express ZEB1. From what I have seen in Uniprot.org, ZEB1 is expressed in myometrium, brain thalamus, cervix carcinoma, leukemic t-cell, cervix carcinoma erythroleukemia, among other tissues. Regarding which tissues have ZEB1 expression, here are a few articles citing expression in various tissues:
Brain, and Thalamus, Pubmed ID: 14702039
Cervix carcinoma, Pubmed ID: 17081983, 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Leukemic T-cell, Pubmed ID: 19690332
Boster Scientific Support
Answered: 2017-09-21
Question
We ordered your anti-AREB6/ZEB1 antibody for IF on brain thalamus in the past. I am using human, and I plan to use the antibody for Flow Cytometry next. I would like examining brain thalamus as well as cervix carcinoma in our next experiment. Do you have any suggestion on which antibody would work the best for Flow Cytometry?
Verified Customer
Verified customer
Asked: 2017-07-24
Answer
I looked at the website and datasheets of our anti-AREB6/ZEB1 antibody and it appears that A00548-2 has been tested on human in both IF and Flow Cytometry. Thus A00548-2 should work for your application. Our Boster satisfaction guarantee will cover this product for Flow Cytometry in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for Flow Cytometry detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2017-07-24
Question
We are currently using anti-AREB6/ZEB1 antibody A00548-2 for human tissue, and we are well pleased with the IHC-P results. The species of reactivity given in the datasheet says human. Is it likely that the antibody can work on zebrafish tissues as well?
Verified Customer
Verified customer
Asked: 2017-06-09
Answer
The anti-AREB6/ZEB1 antibody (A00548-2) has not been validated for cross reactivity specifically with zebrafish tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in zebrafish you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-06-09
Question
My team were satisfied with the WB result of your anti-AREB6/ZEB1 antibody. However we have been able to see positive staining in brain thalamus nucleus using this antibody. Is that expected? Could you tell me where is ZEB1 supposed to be expressed?
R. Li
Verified customer
Asked: 2015-10-19
Answer
From literature, brain thalamus does express ZEB1. Generally ZEB1 expresses in nucleus. Regarding which tissues have ZEB1 expression, here are a few articles citing expression in various tissues:
Brain, and Thalamus, Pubmed ID: 14702039
Cervix carcinoma, Pubmed ID: 17081983, 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Leukemic T-cell, Pubmed ID: 19690332
Boster Scientific Support
Answered: 2015-10-19
