Anti-PCNA antibody

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (AB18197)

ab18197 staining PCNA in tissue sections of the marmoset spleen by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 1% B.S.A. for 10 minutes at 21°C followed by incubation with the primary antibody for 2 hours at 1/6000. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

Image courtesy of Carl Hobbs, Kings College London, U.K.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody (AB18197)

ab18197 staining PCNA in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab18197 at 0.1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PCNA antibody (AB18197)

Overlay histogram showing HeLa cells stained with ab18197 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 20 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18197, 0.05μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab171870, 0.05μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (AB18197)

ab18197 staining PCNA in tissue sections of the cow spleen by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 1% B.S.A. for 10 minutes at 21°C followed by incubation with the primary antibody for 2 hours at 1/4000. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

Image courtesy of Carl Hobbs, King College London, U.K.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (AB18197)

ab18197 staining PCNA in tissue sections of the mouse brain by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 1% B.S.A. for 10 minutes at 21°C followed by incubation with the primary antibody for 2 hours at 1/6000. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

Image courtesy of Carl Hobbs, Kings College London, U.K.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (AB18197)

ab18197 staining PCNA in tissue sections of the rat brain by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 1% B.S.A. for 10 minutes at 21°C followed by incubation with the primary antibody for 2 hours at 1/10000. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

Image courtesy of Carl Hobbs, Kings College London, U.K.

• WB

Project

Western blot - Anti-PCNA antibody (AB18197)

All lanes:

Western blot - Anti-PCNA antibody (ab18197) at 1 µg/mL

Lane 1:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

Lane 2:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg

Lane 3:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg

Lane 4:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg with Human PCNA peptide (<a href='/en-us/products/unavailable/human-pcna-peptide-ab18602'>ab18602</a>)

Lane 5:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg with Human PCNA peptide (<a href='/en-us/products/unavailable/human-pcna-peptide-ab18602'>ab18602</a>)

Lane 6:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg with Human PCNA peptide (<a href='/en-us/products/unavailable/human-pcna-peptide-ab18602'>ab18602</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 29 kDa

Observed band size: 29 kDa,48 kDa,52 kDa

true

Exposure time: 1min

• IHC-P

AbReview39111****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (AB18197)

ab18197 staining PCNA in monkey COS cell pellet by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 1% B.S.A. for 10 minutes at 21°C followed by incubation with the primary antibody for 2 hours at 1/4000. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

Image Courtesy of Carl Hobbs, Kings College London, U.K.

• IHC-P

AbReview39527****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (AB18197)

ab18197 staining PCNA in tissue sections of the goat spleen by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 1% B.S.A. for 10 minutes at 21°C followed by incubation with the primary antibody for 2 hours at 1/4000. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

Image courtesy of Carl Hobbs, Kings College London, U.K.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (AB18197)

ab18197 staining PCNA in tissue sections of the sheep spleen by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 1% B.S.A. for 10 minutes at 21°C followed by incubation with the primary antibody for 2 hours at 1/6000. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

Image courtesy of Carl Hobbs, King College London, U.K.

• WB

Unknown

Western blot - Anti-PCNA antibody (AB18197)

All lanes:

Western blot - Anti-PCNA antibody (ab18197) at 1 µg/mL

All lanes:

Western blot - Recombinant Human PCNA protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pcna-protein-ab85651'>ab85651</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 29 kDa

true

Exposure time: 10s

• WB

Unknown

Western blot - Anti-PCNA antibody (AB18197)

All lanes:

Western blot - Anti-PCNA antibody (ab18197) at 1 µg/mL

All lanes:

Western blot - Recombinant Human PCNA protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pcna-protein-ab85651'>ab85651</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 29 kDa

true

Exposure time: 10s