Immunofluorescent analysis of PCNA (purple) in HeLa (left panel) and NCCIT (right panel) cells. The cells were fixed and permeabilized with ice-cold methanol for 15 minutes, and blocked with 1% Blocker BSA in PBS (Product # 37525) for 15 minutes at room temperature. Cells were stained with a PCNA polyclonal antibody (Product # PA5-16797) at a dilution of 1:50 in 1% Blocker BSA in PBS (Product # 37525) for 1 hour at room temperature, and then incubated with a DyLight 594-conjugated goat anti-rabbit IgG secondary antibody (Product # 35560) at a dilution of 1:250 for 30 minutes at room temperature. Actin (green) was stained with a DyLight 488-conjugated beta-actin monoclonal antibody (Product # MA5-15739-D488) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Dog, Human, Mouse, Non-human primate, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant human PCNA protein|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:50-1:100|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Western Blot (WB)||1:50-1:200|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Methanol fixation is recommended for IF applications. Paraformaldehyde fixation for IF will result in mostly nuclear staining with some cytoplasmic staining.
Expression of proliferating cell nuclear antigen (PCNA) or cyclin or polymerase delta auxiliary protein is elevated in the nucleus during late G1 phase immediately before the onset of DNA synthesis, becoming maximal during S-phase and declining during G2 and M phases. PCNA/cyclin may act as an auxiliary protein of DNA polymerase-delta to play a fundamental role in the initiation of cell proliferation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.