Revision 5

#35031Store 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-P, ChIP, ChIP-seq

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

65 to 75

Source/Isotype:

Rabbit IgG

UniProt ID:

#P15336

Entrez-Gene Id:

1386

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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:400
Chromatin IP 1:50
Chromatin IP-seq 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.

For a carrier free (BSA and azide free) version of this product see product #44072.

Specificity / Sensitivity

ATF-2 (D4L2X) XP® Rabbit mAb recognizes endogenous levels of total ATF-2 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln170 of human ATF-2 protein.

Background

The transcription factor ATF-2 (also called CRE-BP1) binds to both AP-1 and CRE DNA response elements and is a member of the ATF/CREB family of leucine zipper proteins (1). ATF-2 interacts with a variety of viral oncoproteins and cellular tumor suppressors and is a target of the SAPK/JNK and p38 MAP kinase signaling pathways (2-4). Various forms of cellular stress, including genotoxic agents, inflammatory cytokines, and UV irradiation, stimulate the transcriptional activity of ATF-2. Cellular stress activates ATF-2 by phosphorylation of Thr69 and Thr71 (2-4). Both SAPK and p38 MAPK have been shown to phosphorylate ATF-2 at these sites in vitro and in cells transfected with ATF-2. Mutations of these sites result in the loss of stress-induced transcription by ATF-2 (2-4). In addition, mutations at these sites reduce the ability of E1A and Rb to stimulate gene expression via ATF-2 (2).

  1. Abdel-Hafiz, H.A. et al. (1992) Mol Endocrinol 6, 2079-89.
  2. Gupta, S. et al. (1995) Science 267, 389-93.
  3. van Dam, H. et al. (1995) EMBO J 14, 1798-811.
  4. Livingstone, C. et al. (1995) EMBO J 14, 1785-97.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

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
#35031

ATF-2 (D4L2X) XP® Rabbit mAb

Western Blotting Image 1: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from HeLa cells, mock transfected (-) or transfected with SignalSilence® ATF-2 siRNA #6443 (+), using ATF-2 (D4L2X) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using ATF-2 (D4L2X) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
No image available
Immunohistochemistry Image 1: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded Molt-4 cell pellet (left, high) or SKBR3 cell pellet (right, low) using ATF-2 (D4L2X) XP® Rabbit mAb.
Immunohistochemistry Image 2: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using ATF-2 (D4L2X) XP® Rabbit mAb.
Immunohistochemistry Image 3: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using ATF-2 (D4L2X) XP® Rabbit mAb.
Immunohistochemistry Image 4: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using ATF-2 (D4L2X) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemistry Image 5: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using ATF-2 (D4L2X) XP® Rabbit mAb.
Chromatin Immunoprecipitation Image 1: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Jurkat cells treated with TPA #4174 (50 ng/ml) and Ionomycin #9995 (200 ng/ml) for 30 min and ATF-2 (D4L2X) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across JUN gene.
Chromatin Immunoprecipitation Image 2: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Jurkat cells treated with TPA #4174 (50 ng/ml) and Ionomycin #9995 (200 ng/ml) for 30 min and ATF-2 (D4L2X) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 1 (upper), including JUN gene (lower).
Chromatin Immunoprecipitation Image 3: ATF-2 (D4L2X) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Jurkat cells treated with TPA #4174 (50 ng/ml) and Ionomycin #9995 (200 ng/ml) for 30 min and either ATF-2 (D4L2X) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human c-Fos Promoter Primers #4663, human TNF alpha promoter 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.