Anti-PCNA antibody (ab18197)
- Datasheet
- References (96)
- Protocols
Overview
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Product nameAnti-PCNA antibody
See all PCNA primary antibodies -
DescriptionRabbit polyclonal to PCNA
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Host speciesRabbit
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Tested applicationsSuitable for: WB, ICC/IF, IHC-P, IHC-FoFr, Flow Cyt, IPmore details
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Species reactivityReacts with: Mouse, Rat, Sheep, Goat, Cow, Human, Monkey, Zebrafish, Common marmoset
Predicted to work with: Dog, Xenopus laevis
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Immunogen
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Positive control
- This antibody gave a positive signal in HEK 293 (Human embryonic kidney cell line), NIH 3T3 and MEF1 (Mouse embryonic fibroblast cell lines) whole cell lysates. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
Applications
Our Abpromise guarantee covers the use of ab18197 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa). | |
| ICC/IF | Use a concentration of 1 µg/ml. | |
| IHC-P | Use at an assay dependent concentration. | |
| IHC-FoFr | Use at an assay dependent concentration. PubMed: 21895533 | |
| Flow Cyt | Use 0.05µg for 106 cells. | |
| IP | Use at an assay dependent concentration. |
Target
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FunctionThis protein is an auxiliary protein of DNA polymerase delta and is involved in the control of eukaryotic DNA replication by increasing the polymerase's processibility during elongation of the leading strand. Induces a robust stimulatory effect on the 3'-5' exonuclease and 3'-phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. Has to be loaded onto DNA in order to be able to stimulate APEX2.
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Sequence similaritiesBelongs to the PCNA family.
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Post-translational
modificationsUpon methyl methanesulfonate-induced DNA damage, mono-ubiquitinated by the UBE2B-RAD18 complex on Lys-164. This induces non-canonical polyubiquitination on Lys-164 through 'Lys-63' linkage of ubiquitin moieties by the E2 complex UBE2N-UBE2V2 and the E3 ligases, HLTF, RNF8 and SHPRH, which is required for DNA repair. 'Lys-63' polyubiquitination prevents genomic instability on DNA damage. Monoubiquitination at Lys-164 also takes place in undamaged proliferating cells, and is mediated by the DCX(DTL) complex, leading to enhance PCNA-dependent translesion DNA synthesis.
Acetylated in response to UV irradiation. Acetylation disrupts interaction with NUDT15 and promotes degradation. -
Cellular localizationNucleus. Forms nuclear foci representing sites of ongoing DNA replication and vary in morphology and number during S phase. Together with APEX2, is redistributed in discrete nuclear foci in presence of oxidative DNA damaging agents.
- Information by UniProt
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Database links
- Entrez Gene: 515499 Cow
- Entrez Gene: 5111 Human
- Entrez Gene: 18538 Mouse
- Entrez Gene: 25737 Rat
- Entrez Gene: 30678 Zebrafish
- Omim: 176740 Human
- SwissProt: Q3ZBW4 Cow
- SwissProt: P12004 Human
see all -
Alternative names
- ATLD2 antibody
- cb16 antibody
- Cyclin antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (ab18197)Image courtesy of Carl Hobbs, Kings College London, U.K.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 21ºC 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (ab18197)Image courtesy of Carl Hobbs, Kings College London, U.K.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 21ºC 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (ab18197)Image courtesy of Carl Hobbs, King College London, U.K.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 21ºC 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (ab18197)Image courtesy of Carl Hobbs, King College London, U.K.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 21ºC 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (ab18197)Image courtesy of Carl Hobbs, Kings College London, U.K.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 21ºC 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (ab18197)Image courtesy of Carl Hobbs, Kings College London, U.K.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 21ºC 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody (ab18197)Image Courtesy of Carl Hobbs, Kings College London, U.K.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 21ºC 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.
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Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody (ab18197)This image is courtesy of an Abreview submitted by Dr Kirk McManusab18197, at a 1/5000 dilution, staining PCNA in assynchronous HeLa cells. Cells were counter-stained with DAPI (red). For more information please refer to Abreview. -
Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody (ab18197)ICC/IF image of ab18197 stained Hela cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18197, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. -
Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody (ab18197)Image courtesy of Hank Farr by Abreview.ab18197 staining PCNA in Zebrafish gastrula embryos by Immunocytochemistry/ Immunofluorescence (wholemount).
Zebrafish embryos were fixed overnight at 4°C when they had reached 60% epiboly. Cells were fixed in formaldehyde, permeabilized using Proteinase K, blocked with 2% goat serum for 2 hours at 20°C and then incubated with ab18197 at a 1/500 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/500 dilution. Cells were post-fixed in PFA for 20 minutes at room temperature after extensive washing of the secondary antibody.
The left panel shows DAPI stained nuclei, the center panel is PCNA staining, and the right panel is the merged image. -
Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody (ab18197)ICC/IF image of ab18197 stained NIH/3T3 cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18197, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Panel A does not show the Alexa Fluor® 488 channel, Panel B shows the specfic nuclear staining by ab18197. -
Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody (ab18197)This image is courtesy of an abreview submitted by Dr Catherine Greenab18197, at a 1/2000 dilution staining PCNA in MRC5 Sv40 transformed fibroblasts. Cells were counterstained with DAPI (blue). For more information please refer to Abreview. -
Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody (ab18197)ab18197 staining PCNA in SK-N-SH cells treated with KN-62 (ab120421), by ICC/IF. Increase in PCNA nuclear expression correlates with increased concentration of KN-62, as described in literature.
The cells were incubated at 37øC for 24h in media containing different concentrations of ab120421 (KN-62) in DMSO, fixed with 100% methanol for 5 minutes at -20øC and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab18197 (1 æg/ml) was performed overnight at 4øC in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
Western blot - Anti-PCNA antibody (ab18197)All lanes : Anti-PCNA antibody (ab18197) at 1 µg/ml
Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with Human PCNA peptide (ab18602) at 1 µg/ml
Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate with Human PCNA peptide (ab18602) at 1 µg/ml
Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate with Human PCNA peptide (ab18602) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 29 kDa
Observed band size: 29 kDa
Additional bands at: 48 kDa, 52 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute -
Immunoprecipitation - Anti-PCNA antibody (ab18197)This image is courtesy of an Abreview submitted by Antibody Solutions Ltdab18197 staining rat PC12 whole cell lysate by Immunoprecipitation.
ab18197 was incubated with the lysate (at a concentration of 5µg/ml) and a Protein A matrix for 12 hours at 4°C to achieve immunoprecipitation. 400µg of protein were present in the lysate input.
Lane order: PCNA IP (lane 1), Control IP (lane 2), Input 5% (lane 3)
This antibody immunoprecipitates a product of ~34 kDa which is consistent with the mobility of PCNA on SDS-PAGE
ab18197 was also used for the western blot step, at a contration of 1µg/ml
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Western blot - Anti-PCNA antibody (ab18197) -
Western blot - Anti-PCNA antibody (ab18197)
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Flow Cytometry - 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 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22ºC. 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.
Protocols
References
This product has been referenced in:
- Briggs EM et al. Long interspersed nuclear element-1 expression and retrotransposition in prostate cancer cells. Mob DNA 9:1 (2018). Read more (PubMed: 29308092) »
- Zhu L et al. TIPE2 suppresses progression and tumorigenesis of esophageal carcinoma via inhibition of the Wnt/ß-catenin pathway. J Transl Med 16:7 (2018). Read more (PubMed: 29343267) »
