This antibody was tested in IHC using a 48-hpf zebrafish embryo tail. Fluorescence detection was done with HiLyte Fluor™ 488 labeled goat anti-rabbit IgG. Upon addition of the immunizing peptide, the signal was abrogated. Fluorescent western blot analysis using HiLyte Fluor™ 750 shows an immunoreactive band 53 kD, this band was also abrogated upon addition of the immunizing peptide.
Anti-p53, Human or Mouse Specific Antibodies
One of the most important mammalian cell cycle checkpoint proteins is the tumor suppressor protein, p53. In normal, undamaged cells, p53 is rapidly degraded. However, when cells are treated with DNA damage-inducing agents, there is a transient accumulation of p53 protein and it is activated as a transcription factor. In several types of human cancers, p53 is mutated.1,2 Human p53 protein has been shown to be phosphorylated at several N-terminal and C-terminal sites that affect site-specific DNA binding and interaction with other cellular and viral proteins in vitro.3-8 Phosphorylation at serines 6, 9, 15, 20, 33, 37 occurs after cells are exposed either to ionizing radiation or to UV light.9,10 Serines 6 and 15 were demonstrated to be among the strongest and earliest phosphorylated sites in response to DNA damage-induced posttranslational modifications.11,12 AnaSpec’s collection of p53 antibody products includes both phosphospecific and non-phosphospecific solutions. Shown below are western blots of hydroxyurea-treated Cos-7 cells expressing an increasing amount of phosphorylated serine p53’s.
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2. Albrechtsen, N. et al. Oncogene 18, 7706 (1999).
3. Wang, L. et al. J. Biol Chem. 276, 43604 (2001).
4. Xirodimas, D. et al. Oncogene 20, 4972 (2001).
5. Backlund, MG. et al. Cancer Res. 61, 6577 (2001).
6. Sakaguchi, K. et al. J. Biol. Chem. 275, 9278 (2000).
7. Banin, S. et al. Science 281, 1674 (1998).
8. Canman, CE. et al. Science 281, 1677 (1998).
9. Burns, TF. and El-Deiry, WS. J. cell Physiol. 181, 231 (1999).
10. Oren, M. et al. J. Biol. Chem 274, 36031 (1999).
11. Lakin, ND. et al Oncogene 18, 7644 (1999).
12. Higashimoto, Y. et al. J. Biol. Chem. 275, 23199 (2000).
AnaSpec (anaspec.com) is a leading provider of integrated proteomics solutions to the world’s largest biotech, pharmaceutical, and academic research institutions. With a vision for innovation through synergy, AnaSpec focuses on three core technologies: peptides, detection reagents, and antibodies.