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ZEB1 Antibody - N-terminal region (ARP30976_P050)

100 ul
$319.00
In Stock
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Conjugation Options

ARP30976_P050-FITC Conjugated

ARP30976_P050-HRP Conjugated

ARP30976_P050-Biotin Conjugated

Gene Symbol:
ZEB1
Official Gene Full Name:
Zinc finger E-box binding homeobox 1
NCBI Gene Id:
6935
Protein Name:
Zinc finger E-box-binding homeobox 1
Swissprot Id:
P37275
Protein Accession #:
NP_110378
Nucleotide Accession #:
NM_030751
Alias Symbols:
AREB6, BZP, DELTA-EF1, FECD6, MGC133261, NIL-2-A, NIL-2A, NIL2A, TCF8, ZEB, ZFHEP, ZFHX1A, PPCD3, DELTAEF1
Replacement Item:
This antibody may replace item sc-10570 from Santa Cruz Biotechnology.
Description of Target:
ZEB1 is a zinc finger transcription factor that represses T-lymphocyte-specific IL2 gene expression by binding to a negative regulatory domain 100 nucleotides 5-prime of the IL2 transcription start site.
Protein Size (# AA):
1124
Molecular Weight:
124kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Affinity Purified
Application:
IHC, WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express ZEB1.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express ZEB1.
Immunogen:
The immunogen is a synthetic peptide directed towards the N terminal region of human ZEB1
Predicted Species Reactivity:
Cow, Dog, Human, Mouse, Pig, Rabbit, Rat, Zebrafish
Tested Species Reactivity:
Human
Predicted Homology Based on Immunogen Sequence:
Cow: 100%; Dog: 100%; Human: 100%; Mouse: 93%; Pig: 100%; Rabbit: 100%; Rat: 93%; Zebrafish: 100%
Complete computational species homology data:
Anti-ZEB1 (ARP30976_P050)
Peptide Sequence:
Synthetic peptide located within the following region: NPRRNNVTNYNTVVETNSDSDDEDKLHIVEEESVTDAADCEGVPEDDLPT
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Protein Interactions:
SUMO1; CDK6; GTF2A1; UBR4; SUMO2; SIRT1; SOX2; CTBP2; CTBP1; SMAD7; SMAD6; SMARCA4; SERPINH1; SMAD3; EP300; DRAP1; KAT5; SMAD2; SMAD1; CDH1;
Blocking Peptide:
For anti-ZEB1 (ARP30976_P050) antibody is Catalog # AAPP34123
Datasheets/Manuals:
Printable datasheet for anti-ZEB1 (ARP30976_P050) antibody
Publications:

Kothandaraman, N. et al. E2F5 status significantly improves malignancy diagnosis of epithelial ovarian cancer. BMC Cancer 10, 64 (2010). IHC, WB, Cow, Dog, Human, Mouse, Pig, Rabbit, Rat, Zebrafish 20181230

Li, J. et al. A strategy to rapidly identify the functional targets of microRNAs by combining bioinformatics and mRNA cytoplasmic/nucleic ratios in culture cells. FEBS Lett. 584, 3198-202 (2010). IHC, WB, Cow, Dog, Human, Mouse, Pig, Rabbit, Rat, Zebrafish 20547158

Product Reviews

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2 Item(s)

02/01/2017 14:23
  • Overall Experience:
  • Quality:
Product Review: ZEB1 antibody-N-terminal region (ARP30976_P050) in Human Pancreatic cancer cell lines HPAF1 and Capan1 using Western Blot

Product Page for ZEB1 antibody-N-terminal region (ARP30976_P050)
 Researcher: Dr. Pankaj Singh, UNMC, Omaha, NE
 Application: Western blotting
 Species + Tissue/Cell type: Human Pancreatic cancer cell lines HPAF1 and Capan1
How many ug's of tissue/cell lysate run on the gel:
1: 45ug capan1 cell lysate
2: 45 ug HPAF cell lysate
Primary antibody dilution: 1:1000
 Secondary antibody: Anti-Rabbit HRP
Secondary antibody dilution: 1:5000


 Questionnaire:
 

How do Aviva's reagents play a role in your experimental goals?
Used in different biochemical characterization.
How would you rate this antibody on a scale from 1-5 (5=best) and why?
 5/5, Giving specific band.
Would you use this antibody in future experiments? Yes.
Have you used another antibody which has worked in your application? No.
Do you believe the information about the reagent on Aviva's website is correct?
Almost correct.
If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?
We are planning to use it for some of the future experiments
How did you store the antibody after re-suspension? At -20C in small aliquots.

Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):
Human, Pancreatic cancer cell lines: HPAF1 and Capan1, 45 ug
How many different experimental trials were conducted using the antibody sample? 1.
How was this sample prepared?
Cells were lysed in RIPA buffer.
Primary antibody dilution and incubation time:
1:1000, Overnight at 4C.
Secondary antibody used and dilution and incubation time:
1:5000, 2 hour, RT.
 What controls were used in your experiment (positive/negative)?
Beta-tubulin.
Please include your detailed WB Procedure/Protocol here:
_x000D_ IMMUNOBLOTTING PROTOCOL
_x000D_ 1- Prepared the 10 % SDS-PAGE gel.
_x000D_ 2- Loaded 45 ug of total protein (Cell lysate) along with molecular weight markers.
_x000D_ 3- Run the gel initially at 70V, till the dye reached up to the separating gel and then at 110V till the time dye reached to bottom.
_x000D_ 4- Cut PVDF membrane to the same size of the gel. Soaked the membrane in 100% Methanol for 10sec then in water for 20 sec and finally 20 min in transfer buffer.
_x000D_ 5- Arranged the Electro-transfer apparatus with gel-membrane sandwich and connected to the power supply at constant Amp (225 mA) for 1hour.
_x000D_ 6- Disconnected the transfer apparatus and transferred the membrane in blocking solution [5% Skimmed in PBST (0.1%)] for 1 hour.
_x000D_ 7- After blocking transferred the membrane in Primary antibody solution (1:1000 dilutions in PBST) and incubated at 4C for over-night.
_x000D_ 8- Washed the membrane 3 times with PBST (0.1%) for 10 minutes each.
_x000D_ 9- Incubated the membrane with respective secondary antibody (1:5000 dilutions in PBST) for 2 hour at room temperature.
_x000D_ 10- Washed the membrane 3 times with PBST (0.1%) for 10 minutes each.
_x000D_ 11- Developed the blots using standard protocol.
_x000D_
_x000D_

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02/01/2017 14:23
  • Overall Experience:
  • Quality:
Product Review: ZEB1 antibody-N-terminal region (ARP30976_P050) in Human Capan1 cells (Pancreatic cancer cell line) using IHC

Product Page for ZEB1 antibody-N-terminal region (ARP30976_P050)
Researcher: Dr. Pankaj Singh, UNMC, Omaha, NE
Application: IHC
Species + Tissue/Cell type: Human Capan1 cells (Pancreatic cancer cell line)
How many ug's of tissue/cell lysate run on the gel:
Human Capan1 cells (Pancreatic cancer cell line)
Primary antibody dilution: 1:300
Secondary antibody: Anti-rabbit-AlexaFluor-488
Secondary antibody dilution: 1:200

Questionnaire:
How do Aviva's reagents play a role in your experimental goals?
Used in different biochemical characterization.

How would you rate this antibody on a scale from 1-5 (5=best) and why?
5/5, Giving specific band.

Would you use this antibody in future experiments?
Yes.

Have you used another antibody which has worked in your application?
No.

Do you believe the information about the reagent on Aviva's website is correct?
Most of the time its correct.

If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?
We are planning to use it for some of the future experiments.

_x000D_ How did you store the antibody after re-suspension?
_x000D_ At -20C in small aliquots.
_x000D_
_x000D_ Sample Description (please include species type and tissue/cell type):
_x000D_ Pancreatic cancer cell line Capan1.
_x000D_
_x000D_ Please explain fixation solution/method used (formalin, periodate-lysine-paraformaldehyde, acetone, etc.)?
_x000D_ With 4% Paraformaldehyde.
_x000D_
_x000D_ How many different experimental trials were conducted using the antibody sample?
_x000D_ 1.
_x000D_
_x000D_ Primary antibody dilution, incubation time and temperature:
_x000D_ 1:300, 2 hour, RT.
_x000D_
_x000D_ Secondary antibody used, dilution, incubation time and temperature:
_x000D_ 1: 200, 1:30 hour, RT.
_x000D_
_x000D_ From your IHC/ICC images, briefly explain the colors of each stain and counterstain:
_x000D_ Green- Alexa flour-488, Blue-DAPI.
_x000D_
_x000D_ Did you use an antigen retrieval method? If so, please explain?
_x000D_ No.
_x000D_
_x000D_ What controls were used in your experiment?
_x000D_ Secondary Antibody control.
_x000D_
_x000D_ Please include your detailed tissue preparation and staining procedure/protocol here:
_x000D_ 1. Seeded 250,000 cells on coverslips in 12 well plate and grown over night
_x000D_ 2. Washed the cells with 1X PBS.
_x000D_ 3. Fixed the cell in 4% Paraformaldehyde for 15 minute at RT.
_x000D_ 4. Incubated the cells with 0.1M Glycine solution for 15 minute at RT to quench the excess paraformaldehyde.
_x000D_ 5. Washed the cells with 1X PBS twice and left them in 1% BSA in PBS for over-night.
_x000D_ 6. Incubated the cells in 0.1% Triton-X 100 in PBS for 10 minute to permeabilize the cells.
_x000D_ 7. Washed the cells with 1X PBS three times.
_x000D_ 8. Incubated the cells with DMEM for 5 minute.
_x000D_ 9. Added 1° Antibody with 1:300 dilutions in 1% BSA in PBST (0.1%) and incubated at RT for 2 hour.
_x000D_ 10. Washed the cells with 1X PBS three times for 5 minutes each.
_x000D_ 11. Added 2° Antibody (Alexa Fluor-488, Jackson Immuno Research Laboratories, Inc) with 1:200 dilution in 1% BSA in PBST(0.1%) and incubated at RT for 1:30 hour.
_x000D_ 12. Washed the cells with 1X PBS three times for 5 minutes each.
_x000D_ 13. Added the Anti-fade with DAPI on glass slides.
_x000D_ 14. Mounted the coverslips (cells facing downward) on the glass slide gently.
_x000D_ 15. Absorbed the extra liquid with tissue paper.
_x000D_ 16. Sealed the edges of coverslips with the help of nail paint.
_x000D_ 17. Stored in the slide box at 4C.
_x000D_ 18. Captured the image.
_x000D_
_x000D_

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