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PCNA Rabbit pAb

sample: A549 cells( (blue) Ice-cold 70% ethanol was added and the cells were incubated for overnight at 4 °C. and then resuspended in ice-cold 90% methanol for 30 min on ice.) Primary Antibody:Rabbit Anti-PCNA antibody(AP93981) (green) Isotype Control Antibody: Rabbit IgG (orange) Secondary Antibody: F(ab’)2 fragment goat anti-rabbit IgG-FITC (white blue)
Blank control (blue line): U251 (blue). Primary Antibody (green line): Rabbit Anti-PCNA antibody (AP93981) Dilution: 3 µg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE Dilution: 1 µg /test. Protocol The cells were fixed with 2% paraformaldehyde (10 min)and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
25 ug total protein per lane of various lysates (see on figure) probed with PCNA polyclonal antibody, unconjugated (AP93981) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (AP93981) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (AP93981) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (AP93981) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Liver Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (AP93981) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.

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BACKGROUND

Product Information
Application Applications Legend: WB=Western Blot IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin-embedded Sections) IHC-F=Immunohistochemistry (Frozen Sections) IF=Immunofluorescence FC=Flow Cytopmetry IC=Immunochemistry ICC=Immunocytochemistry E=ELISA IP=Immunoprecipitation DB=Dot Blot CHIP=Chromatin Immunoprecipitation FA=Fluorescence Assay IEM=Immuno electron microscopy EIA=Enzyme Immunoassay	WB, IHC-P, IHC-F, IF
Reactivity	Human
Host	Rabbit
Clonality	Polyclonal

Additional Information
Dilution	WB=1:500-2000,IHC-P=1:200-800,IHC-F=1:200-800,IF=1:200-800,Flow-Cyt=1 µg/Test
Format	0.01M TBS(pH7.4), 0.09% (W/V) sodium azide and 50% Glyce
Storage	Store at -20 ℃ for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 ℃.

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Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

Background

This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

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Cat# AP93981

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Recombinant Antibody

PCNA Rabbit pAb

PCNA Rabbit pAb

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