Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-ATF-2 (Thr71) Antibody #9221

Applications Reactivity Sensitivity MW (kDa) Source
W IP IHC-P IHC-F IF-IC F H M R Mk Endogenous 70 Rabbit

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

Protocols

Specificity / Sensitivity

Phospho-ATF-2 (Thr71) Antibody detects endogenous levels of ATF-2 only when phosphorylated at threonine 71. This antibody does not cross-react with phosphorylated c-Jun, CREB or other transcription factors. It recognizes both Thr69/Thr71 dually phosphorylated ATF-2 and Thr71 singly phosphorylated ATF-2 equally well.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr71 of human ATF2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, C6 and NIH/3T3 cells, untreated or anisomycin-treated, using Phospho-ATF-2 (Thr71) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma showing nuclear localization, using Phospho-ATF-2 (Thr71) Antibody .

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistchemical analysis of paraffin-embedded human colon carcinoma using Phospho-ATF-2 (Thr71) Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-ATF-2 (Thr71) Antibody.

IHC-F (frozen)

IHC-F (frozen)

Immunhistochemical analysis of frozen H1650 xenograft using Phospho-ATF-2 (Thr71) Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or Anisomycin-treated (green), using Phospho-ATF-2 (Thr71) Antibody compared to a nonspecific negative control antibody (red).


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, either untreated (left) or anisomycin-treated (right), using Phospho-ATF-2 (Thr71) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

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, 2079-2089.
  2. Gupta, S. et al. (1995) Science 267, 389-393.
  3. van Dam, H. et al. (1995) EMBO J. 14, 1798-1811.
  4. Livingstone, C. et al. (1995) EMBO J. 14, 1785-1797.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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