Revision 5

#70512Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-Bond, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

200

Source/Isotype:

Rabbit IgG

UniProt ID:

#P37275

Entrez-Gene Id:

6935

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:200 - 1:800
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #83243.

Specificity / Sensitivity

ZEB1 (E2G6Y) XP® Rabbit mAb recognizes endogenous levels of total ZEB1 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein corresponding to the central region of human ZEB1 protein.

Background

ZEB family proteins are zinc-finger and homeobox domain containing transcription factors. There are two members in mammals, ZEB1 (δ-EF1, TCF8, AREB6) and ZEB2 (SIP1). ZEB1 and ZEB2 contain two separate zinc-finger domains and a homeodomain (1). While ZEB proteins mainly function as transcriptional suppressors, they are able to activate transcription, dependent on DNA-context and cell type (1). One of the targets suppressed by ZEB proteins is E-cadherin. Downregulation of E-cadherin is one of the hallmarks of epithelial mesenchymal transition (EMT), a critical feature of normal embryonic development, which is also utilized by malignant epithelial tumors to spread beyond their origin (2-4). ZEB1 mutations are associated with posterior corneal dystrophy, and ZEB2 mutations were reported to be associated with Hirschsprung (HSCR) disease (5-8).

  1. Vandewalle, C. et al. (2009) Cell Mol Life Sci 66, 773-87.
  2. Aigner, K. et al. (2007) Oncogene 26, 6979-88.
  3. Peinado, H. et al. (2007) Nat Rev Cancer 7, 415-28.
  4. Moreno-Bueno, G. et al. (2008) Oncogene 27, 6958-69.
  5. Krafchak, C.M. et al. (2005) Am J Hum Genet 77, 694-708.
  6. Aldave, A.J. et al. (2007) Am J Med Genet A 143A, 2549-56.
  7. Dastot-Le Moal, F. et al. (2007) Hum Mutat 28, 313-21.
  8. Garavelli, L. and Mainardi, P.C. (2007) Orphanet J Rare Dis 2, 42.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#70512

ZEB1 (E2G6Y) XP® Rabbit mAb

Western Blotting Image 1: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using ZEB1 (E2G6Y) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunoprecipitation Image 1: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunoprecipitation of ZEB1 protein from Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is ZEB1 (E2G6Y) XP® Rabbit mAb. Western blot analysis was performed using ZEB1 (E2G6Y) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunohistochemistry Image 1: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 2: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human mucoepidermoid carcinoma of the larynx using ZEB1 (E2G6Y) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 1: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemistry Image 2: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemistry Image 3: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemistry Image 4: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemistry Image 5: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor using ZEB1 (E2G6Y) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 6: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse lung using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemistry Image 7: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemistry Image 8: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded HT-1080 cell pellet (left, positive) or HCC1806 cell pellet (right, negative) using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunofluorescence Image 1: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HT-1080 cells (left, positive) and HCC1806 cells (right, negative) using ZEB1 (E2G6Y) XP® Rabbit mAb (green), Cytochrome c (6H2.B4) Mouse mAb #12963 (yellow), and Alexa Fluor® 647 Phalloidin #8940 (red). Slides were mounted with ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow Cytometry Image 1: ZEB1 (E2G6Y) XP® Rabbit mAb Expand Image
Flow cytometric analysis of HCC1806 cells (blue, negative) and HT-1080 cells (green, positive) using ZEB1 (E2G6Y) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.