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.

使用に関する制限

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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.