Revision 5

#14069Store 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, IF-IC, ChIP, ChIP-seq, C&R

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

52

Source/Isotype:

Rabbit IgG

UniProt ID:

#P14921

Entrez-Gene Id:

2113

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:2000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:3200 - 1:6400
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:50

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.

Specificity / Sensitivity

ETS-1 (D8O8A) Rabbit mAb recognizes endogenous levels of total ETS-1 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser179 of human ETS-1 protein.

Background

ETS-1 is a proto-oncoprotein that belongs to the E26 Transformation-specific Sequence (ETS) family of transcription factors that share a unique and highly conserved DNA binding domain (1). ETS-1 plays important roles in vascular development and angiogenesis (2), and vascular inflammation and remodeling (3). The target genes of ETS-1 include receptor tyrosine kinases, MMPs, and cell adhesion molecules (4-6). In addition, ETS-1 is involved in regulation of energy metabolism in cancer cells (7). ETS-1 activity is regulated by two different types of phosphorylation sites. While phosphorylation at a cluster of serine residues in the exon VII domain by CaMKII inhibits ETS-1 DNA binding activity (8), phosphorylation at its Thr38 site by Ras activates ETS-1 (9).

  1. Bartel, F.O. et al. (2000) Oncogene 19, 6443-54.
  2. Sato, Y. (2001) Cell Struct Funct 26, 19-24.
  3. Oettgen, P. (2006) Circ Res 99, 1159-66.
  4. Dube, A. et al. (1999) Circ Res 84, 1177-85.
  5. Lelièvre, E. et al. (2000) Oncogene 19, 2438-46.
  6. Ghosh, S. et al. (2012) J Biol Chem 287, 15001-15.
  7. Verschoor, M.L. et al. (2010) PLoS One 5, e13565.
  8. Pognonec, P. et al. (1988) EMBO J 7, 977-83.
  9. Yang, B.S. et al. (1996) Mol Cell Biol 16, 538-47.

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 IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

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.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
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
#14069

ETS-1 (D8O8A) Rabbit mAb

Western Blotting Image 1: ETS-1 (D8O8A) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using ETS-1 (D8O8A) Rabbit mAb.
Immunoprecipitation Image 1: ETS-1 (D8O8A) Rabbit mAb Expand Image
Immunoprecipitation of ETS-1 protein from OVCAR8 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ETS-1 (D8O8A) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ETS-1 (D8O8A) Rabbit mAb
Immunofluorescence Image 1: ETS-1 (D8O8A) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of OVCAR8 (high expression, left), A549 (moderate expression, center), and HeLa (low expression, right) cells using ETS-1 (D8O8A) Rabbit mAb (green). Actin filaments were labeled with Dylight 554 Phalloidin #13054 (red).
Chromatin Immunoprecipitation Image 1: ETS-1 (D8O8A) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and ETS-1 (D8O8A) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795.
Chromatin Immunoprecipitation Image 2: ETS-1 (D8O8A) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and ETS-1 (D8O8A) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 1 (upper), including RNF187 (lower).
Chromatin Immunoprecipitation Image 3: ETS-1 (D8O8A) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and either ETS-1 (D8O8A) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human c-Fos Promoter Primers #4663, human c-Myc intron 1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT and RUN Image 1: ETS-1 (D8O8A) Rabbit mAb Expand Image
CUT&RUN was performed with K-562 cells and ETS-1 (D8O8A) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across ATXN7L3, a known target gene of ETS-1 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 2: ETS-1 (D8O8A) Rabbit mAb Expand Image
CUT&RUN was performed with K-562 cells and ETS-1 (D8O8A) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 17 (upper), including ATXN7L3 (lower), a known target gene of ETS-1 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 3: ETS-1 (D8O8A) Rabbit mAb Expand Image
CUT&RUN was performed with K-562 cells, then treated with β-estradiol (10 nM) for 45 minutes and either ETS-1 (D8O8A) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human ATXN7L3 promoter primers, human TMEM53 exon 1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.