Product Pathways - MAPK Signaling
PhosphoPlus® ATF-2 (Thr71) Antibody Kit #9220
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|---|---|
| Phospho-ATF-2 (Thr71) Antibody # 9221 | 100 microliters | W IP IHC-P IHC-F IF-IC F | H M R Mk | 70 | Rabbit |
| ATF-2 (20F1) Rabbit mAb # 9226 | 100 microliters | W IP IHC-P | H M R Mk | 65 to 75 | Rabbit |
| Anti-rabbit IgG, HRP-linked Antibody # 7074 | 50 microliters | Goat | |||
| Anti-biotin, HRP-linked Antibody # 7075 | 100 microliters | Goat | |||
| Biotinylated Protein Ladder Detection Pack # 7727 | 100 microliters | ||||
| 20X LumiGLO® Reagent and 20X Peroxide # 7003 | 5 milliliters | ||||
| ATF-2 Control Cell Extracts # 9223 | 50 microliters |
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
Specificity / Sensitivity
Phospho-ATF-2 (Thr71) Antibody detects endogenous levels of ATF-2 only when phosphorylated at Thr71. It recognizes this site regardless of the phosphorylation state of Thr69. ATF-2 (20F1) Rabbit mAb detects endogenous levels of total ATF-2 protein. Neither antibody cross-reacts with c-Jun, CREB or other transcription factors.
Western Blotting
Western blot analysis of extracts from HeLa, C6 and NIH/3T3 cells, untreated or anisomycin-treated, using Phospho-ATF-2 (Thr71) Antibody #9221.
Western Blotting
Western blot analysis of extracts from 293 and NIH/3T3 cells, untreated or UV-treated, using ATF-2 (20F1) Rabbit mAb #9226.
IHC-P (paraffin)
Immunohistochemical staining of paraffin-embedded human breast carcinoma, using Phospho-ATF-2 (Thr71) Antibody #9221 showing nuclear localization of phosphorylated ATF-2.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma, using ATF-2 (20F1) Rabbit mAb #9226.
IHC-FL (floating)
Confocal images of double immunostaining in the rat hippocampus, control or 24 hours of reperfusion following 15 minute transient cerebral ischemia, using Phospho-ATF-2 (Thr71) Antibody # 9221 (green) and calbindin antibody (red). (Provided by Dr. Bingren Hu, University of Miami School of Medicine, Florida.)
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr71 of human ATF-2 (Phospho-ATF-2 (Thr71) Antibody), or with a synthetic peptide (KLH-coupled) derived from the amino terminal sequence of human ATF-2 (ATF-2 (20F1) Rabbit mAb). Antibodies are purified by protein A and peptide affinity chromatography.
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).
- Abdel-Hafiz, H.A. et al. (1992) Mol. Endocrinol. 6, 2079-2089.
- Gupta, S. et al. (1995) Science 267, 389-393.
- van Dam, H. et al. (1995) EMBO J. 14, 1798-1811.
- Livingstone, C. et al. (1995) EMBO J. 14, 1785-1797.
Application References
- de Ruiter, N.D. et al. (2000) Mol Cell Biol 20, 8480-8. This article references the use of PhosphoPlus® ATF-2 (Thr71) Antibody Kit in the following applications: Western Blotting
- Kacharmina, J.E. et al. (2000) Proc Natl Acad Sci U S A 97, 11545-50. This article references the use of PhosphoPlus® ATF-2 (Thr71) Antibody Kit in the following applications: IC-ABC
- Marinissen, M.J. et al. (2001) Genes Dev 15, 535-53. This article references the use of PhosphoPlus® ATF-2 (Thr71) Antibody Kit in the following applications: Western Blotting
- Ouwens, D.M. et al. (2002) EMBO J 21, 3782-93. This article references the use of PhosphoPlus® ATF-2 (Thr71) Antibody Kit in the following applications: Western Blotting
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Companion Products
- 9260 PhosphoPlus® c-Jun (Ser63) II and c-Jun (Ser73) Antibody Kit
- 9164 Phospho-c-Jun (Ser73) Antibody
- 9210 PhosphoPlus® p38 MAP Kinase (Thr180/Tyr182) Antibody Kit
- 9211 Phospho-p38 MAPK (Thr180/Tyr182) Antibody
- 9230 PhosphoPlus® MKK3/MKK6 (Ser189/207) Antibody Kit
- 9231 Phospho-MKK3/MKK6 (Ser189/207) Antibody
- 9151 Phospho-SEK1/MKK4 (Thr261) Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 9226 ATF-2 (20F1) Rabbit mAb
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
Rabbit Monoclonals Produced UsingTechnology from Epitomics, Inc. UnderU.S. patent No. 5,675,063.
