ATF2 (Phospho-Thr69,71) Transcription Factor Assay Kit

ATF2 (Phospho-Thr^69,71) Transcription Factor Assay Kit

Cayman Chemical Item Number 600130

Activating Transcription Factor 2

Description

ATF2 (Activating Transcription Factor 2) is a sequence-specific DNA-binding protein belonging to the bZIP family of transcription factors that bind with high affinity to the octameric cyclic AMP-responsive element (CRE) in the promoter region of target genes.¹ ATF2 mediates both transcription and DNA damage control through its phosphorylation/activation in response to inflammatory cytokines, UV irradiation, alkylating compounds, and other cellular stressors. The activated Phospho-Thr^69,71 species of ATF2 (pThr69,71 ATF2; phosphorylated at both Thr⁶⁹ and Thr⁷¹) is specifically mediated by JNK/p38.² phospho-Thr^69,71 ATF2 can form homodimers or selective heterodimers with other members of the ATF/CREB family including Fos, Fra2, c-Jun, CREB and ATF1, yielding a diverse array of DNA-binding complexes which contribute to regulatory specificity among bZIP family proteins. Recent studies implicate the ATF/CREB family of transcription factors in key cellular processes including cell cycle control, drug resistance, apoptosis, tumorigenesis, and insulin biosynthesis.³ Cayman’s ATF2 (Phospho-Thr^69,71) Transcription Factor Assay is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts. A 96-well enzyme-linked immunosorbent assay (ELISA) replaces the cumbersome radioactive electrophoretic mobility shift assay (EMSA). A specific double-stranded DNA (dsDNA) sequence containing the cAMP-response element (CRE) is immobilized onto the wells of a 96-well plate. ATF2 contained in a nuclear extract, binds specifically to the CRE. The activated ATF2 transcription factor complex is detected by addition of a specific primary antibody directed against the phospho-Thr^69,71 epitope on ATF2. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm.

1 Hay, C.W., Ferguson, L.A., and Docherty, K. ATF-2 stimulates the human insulin promoter through the conserved CRE2 sequence. Biochim Biophys Acta 1769 79-91 (2007).

2 Vlahopoulos, S.A., Logotheti, S., Mikas, D., et al. The role of ATF-2 in oncogenesis. BioEssays 30 314-327 (2008).

3 Bhoumik, A., Lopez-Bergami, P., and Ronai, Z. ATF2 on the double - activating transcription factor and DNA damage response protein. Pigment Cell Res 20 498-506 (2007).

Synonyms

Activating Transcription Factor 2

Stability

6 months

Storage

-80°C

Shipping

Dry ice in continental US; may vary elsewhere

Background Reading

Hay, C.W., Ferguson, L.A., and Docherty, K. ATF-2 stimulates the human insulin promoter through the conserved CRE2 sequence. Biochim Biophys Acta 1769 79-91 (2007).

Vlahopoulos, S.A., Logotheti, S., Mikas, D., et al. The role of ATF-2 in oncogenesis. BioEssays 30 314-327 (2008).

Bhoumik, A., Lopez-Bergami, P., and Ronai, Z. ATF2 on the double - activating transcription factor and DNA damage response protein. Pigment Cell Res 20 498-506 (2007).

600130-96well

Transcription Factor Binding Assay Buffer (4X)

1 ea

Transcription Factor Antibody Binding Buffer (10X)

1 ea

Transcription Factor Developing Solution

1 ea

Transcription Factor Stop Solution

1 ea

Transcription Factor Reagent A

1 ea

Transcription Factor Goat Anti-Mouse HRP Conjugate