Cell Signaling Technology

Product Pathways - Metabolism

ATF-4 (D4B8) Rabbit mAb #11815

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H (M) (R) Endogenous 49 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

ATF-4 (D4B8) Rabbit mAb recognizes endogenous levels of total ATF-4 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human ATF-4 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 and HeLa cells, untreated (-) or tunicamycin-treated (2 μg/ml, 8 hr; +), using ATF-4 (D4B8) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

IP

IP

Immunoprecipitation of ATF-4 from extracts of 293 cells, treated with tunicamycin (2 μg/ml, 8 hr), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ATF-4 (D4B8) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ATF-4 (D4B8) Rabbit mAb. Mouse Anti-rabbit IgG (Light-Chain Specific) (L57A3) mAb #3677 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as secondary antibodies.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or tunicamycin-treated (2 μg/ml, 8 hr; right), using ATF-4 (D4B8) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).


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.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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