Revision 7

#97038Store 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, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

49

Source/Isotype:

Mouse IgG1 kappa

UniProt ID:

#P18848

Entrez-Gene Id:

468

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

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

ATF-4 (E4Q4E) Mouse mAb recognizes endogenous levels of total ATF-4 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro134 of human ATF-4 protein.

Background

ATF-4, an activating transcription factor/cAMP-response element-binding protein family member, functions in the PERK and eIF2α ER stress responsive pathway (1-3). ER stress represses the translation of the majority of mRNAs, but selectively stimulates the translation of certain mRNAs including that of ATF-4 (2). Induced expression of ATF-4 increases the expression of genes critical for the recovery from ER stress (4).

  1. Fawcett, T.W. et al. (1999) Biochem J 339 (Pt 1), 135-41.
  2. Harding, H.P. et al. (2000) Mol Cell 6, 1099-108.
  3. van Huizen, R. et al. (2003) J Biol Chem 278, 15558-64.
  4. Yusta, B. et al. (2006) Cell Metab 4, 391-406.

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 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.
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 7
#97038

ATF-4 (E4Q4E) Mouse mAb

Western Blotting Image 1: ATF-4 (E4Q4E) Mouse mAb Expand Image
Western blot analysis of extracts from 293T cells, vehicle-treated (-) or treated with Tunicamycin #12819 (2 μg/ml, 8 hr; +), and HeLa cells, vehicle-treated (-) or treated with Tunicamycin #12819 (2 μg/ml, 7 hr; +), using ATF-4 (E4Q4E) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunofluorescence Image 1: ATF-4 (E4Q4E) Mouse mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells, vehicle-treated (left) or treated with Tunicamycin #12819 (2 µg/ml, 7 hr; right), using ATF-4 (E4Q4E) Mouse mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
Flow Cytometry Image 1: ATF-4 (E4Q4E) Mouse mAb Expand Image
Flow cytometric analysis of HeLa cells (untreated, blue) or treated with Tunicamycin (2 µg/mL for 8 hr, green) using ATF-4 (E4Q4E) Mouse mAb (solid line) or concentration-matched Mouse (G3A1) mAb IgG1 XP® Isotype Control #5415 (dashed line). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.