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Safety Data Sheet (SDS) Purified Anti-human/mouse/rat PCNA Antibody     Product Data Sheet (PDF)    
Purified Anti-human/mouse/rat PCNA Antibody
2139505 25 µg $55.00       
2139510 100 µg $100.00       
Clone: PC10
Isotype: Mouse IgG2a, κ
Reactivity: Human, Mouse, Rat, Cross-Reactivity: Other species
Immunogen: Recombinant rat PCNA
Formulation: Phosphate-buffered solution, pH7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application:

ICFC - Quality tested
IF, IHC - Validated
IP, WB - Reported in the literature

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.
Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent intracellular staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤0.125 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

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 HYE, et al. 2006. J. Biol. Chem. 10:1074.
10. Fulvio MD, et al. 2006. Oncogene 25:3932.
11. Eswarakumar VP and Schlessinger J. 2007. Proc. Natl. Acad. Sci. USA 104:3937.
12. Henkels KM, et al. 2009. Biochem Biophys Res Commun. 389:224. PubMed
13. Brobeli A, et al. 2010. Blood Cells Mol Dis. 45:159. PubMed
14. Wallace HA, et al. 2014. Development. 141:1332. PubMed
15. Mizokami A, et al. 2014. Bone. 69:68. PubMed

Hela cell nuclear extract was

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.

HeLa cells were fixed with

HeLa cells were fixed with 1% paraformaldehyde (PFA) for 10 minutes, permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 30 minutes. Then the cells were intracellularly stained with 5 µg/ml of purified anti-human/mouse/rat PCNA (clone PC10) in blocking buffer overnight at 4°C, followed by DyLight™ 594 anti-mouse IgG (red) and Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes. Nuclei were counterstained with DAPI (blue). The image was captured with 20X objective.



Description:

The PC10 monoclonal antibody reacts with proliferating cell nuclear antigen also known as PCNA or the DNA polymerase δ auxiliary protein. PCNA 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.

Other Names: Proliferating Cell Nuclear Antigen, DNA Polymerase δ Auxiliary Protein
Structure: DNA-polymerase sliding clamp, trimeric ring; 36 kD
Distribution: Nuclear, all proliferating cells
Function: RAD6-dependent DNA repair pathway; increases DNA polymerase δ processibility during elongation of the leading strand
Ligand Receptor: 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.