Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis and/or Western blotting. For immunofluorescent staining, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µl volume or 100 µl of whole blood. For Western blotting, the suggested use of this reagent is ≤ 1.0 µg/ml. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
Additional reported applications (for the relevant formats) include: immunohistochemical staining2,5,6 of acetone-fixed frozen sections and formalin-fixed paraffin-embedded tissue sections, immunoprecipitation, immunofluorescence microscopy9, and Western blotting10.
Application References:
1. Ogata, K., et al., 1985. J. Immunol. 135:2623. 2. Garcia, R., et al., 1989. Am. J. Pathol. 134:733. 3. Landberg, G., et al., 1990. Exp. Cell. Res. 187:111. 4. Waseem, N., et al., 1990. J. Cell Sci. 96:121. 5. Yu, C., et al., 1991. Histopathology 19:29. 6. Wilkins, B., et al., 1992. J. Pathol. 166:45. 7. Yang, W., et al., 1996. Human Pathol. 27:70. 8. Galkowska, H., et al., 1996. Vet. Immunol. Immunopathol. 53:329. 9. Chou, H-Y. E., et al., 2006. J. Biol. Chem. 10:1074. 10. Fulvio, M. D., et al., 2006. Oncogene 25:3932. 11. Eswarakumar, V. P. and Schlessinger, J., 2007. Proc. Natl. Acad. Sci. USA 104:3937.PubMed 12. Chou, HY.,et al.2006.J Biol Chem.281:15201. PubMed 13. Di Fulvio, M.,et al.2006. Oncogene.25:3032.PubMed
Hela cell nuclear extract was resolved by electrophoresis, transferred to nitrocellulose and probed with monoclonal anti-PCNA antibody. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system.
Description:
PCNA, also known as DNA polymerase δ auxiliary protein, is a 36 kD trimeric ring that acts as a DNA-polymerase sliding clamp expressed in the nucleus of all proliferating cells. A prime function of PCNA appears to be increasing DNA polymerase δ processibility during elongation of the leading strand. PCNA is a useful marker for DNA synthesis and is highly conserved among most species, thus highlighting the very broad reactivity of this antibody. The PC10 monoclonal antibody recognizes human and mouse PCNA (cross-reactive with other species) and has been shown to be useful for flow cytometry, immunohistochemistry (frozen and formalin-fixed, paraffin-embedded tissues), immunoprecipitation, Western blotting, and immunofluorescence staining.
Other Names:
Proliferating Cell Nuclear Antigen, DNA Polymerase δ Auxiliary Protein
RAD6-dependent DNA repair pathway; increases DNA polymerase δ processibility during elongation of the leading strand
Modification:
Ubiquitination, Sumoylation
Interaction:
PCNA, DNA polymerase δ, Rad6, Rad18, UBC9, MMS2, UBC13, RAD5
Antigen References:
1. Travali, S., et al., 1989. J. Biol. Chem. 264:7466. 2. Waseem, N., et al., 1990. J. Cell Sci. 96:121. 3. Hall, P., et al., 1990. J. Pathol. 162:285. 4. Landberg, G., et al., 1991. Cancer Res. 51:4570. 5. Woods A., et al., 1991. Histopathol. 19:21. 6. Hoege, C., et al., 2002. Nature 419:135. 7. Yue, H., et al., 2003. World J. Gastroenterol. 9:377. 8. Shan, B., et al., 2003. J. Biol. Chem. 278:44009.
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