製品の概要

  • 製品名Anti-PCNA antibody
    PCNA 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to PCNA
  • アプリケーション適用あり: WB, ICC/IF, IHC-P, IHC-FoFr, Flow Cyt, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Sheep, Goat, Cow, Human, Monkey, Zebrafish, Marmoset (common)
    交差が予測される動物種: Dog, Xenopus laevis
  • 免疫原

    Synthetic peptide corresponding to Human PCNA aa 200 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P12004
    (Peptide available as ab18602, ab2427)

  • ポジティブ・コントロール
    • 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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能This 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.
  • 配列類似性Belongs to the PCNA family.
  • 翻訳後修飾Upon 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.
  • 細胞内局在Nucleus. 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
  • 参照データベース
  • 別名
    • ATLD2 antibody
    • cb16 antibody
    • Cyclin antibody
    • DNA polymerase delta auxiliary protein antibody
    • etID36690.10 antibody
    • fa28e03 antibody
    • fb36g03 antibody
    • HGCN8729 antibody
    • MGC8367 antibody
    • Mutagen-sensitive 209 protein antibody
    • OTTHUMP00000030189 antibody
    • OTTHUMP00000030190 antibody
    • PCNA antibody
    • Pcna/cyclin antibody
    • PCNA_HUMAN antibody
    • PCNAR antibody
    • Polymerase delta accessory protein antibody
    • Proliferating cell nuclear antigen antibody
    • wu:fa28e03 antibody
    • wu:fb36g03 antibody
    see all

Anti-PCNA antibody 画像

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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.

    See Abreview

  • ab18197, 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.

    See Abreview

  • 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.

  • 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.

    See Abreview

  • 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.
  • ab18197, 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.

    See Abreview

  • 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.
  • 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
    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
  • ab18197 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

    See Abreview


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 29 kDa

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 29 kDa


    Exposure time : 10 seconds
  • 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.

Anti-PCNA antibody (ab18197) 使用論文

This product has been referenced in:
  • Tian LQ  et al. MicroRNA-197 inhibits cell proliferation by targeting GAB2 in glioblastoma. Mol Med Rep N/A:N/A (2016). Read more (PubMed: 27035789) »
  • Deng M  et al. Silencing cyclin-dependent kinase inhibitor 3 inhibits the migration of breast cancer cell lines. Mol Med Rep 14:1523-30 (2016). WB ; Human . Read more (PubMed: 27314680) »

See all 26 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Dog Cell lysate - whole cell (MDCK)
Gel Running Conditions Non-reduced Non-Denaturing (Native) (4-12%)
Loading amount 15 µg
Specification MDCK
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Nov 25 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Pig Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 25 µg
Specification Intestinal tissue
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

投稿 Jul 08 2016

Abreviews
Application Western blot
Sample Human Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

投稿 Jul 08 2016

Application Western blot
Sample Rat Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

投稿 Jul 08 2016

Application Western blot
Sample Mouse Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount 14 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

投稿 Jul 08 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (lymphoma)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification lymphoma
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
Username

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投稿 Dec 17 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (prostate cancer cell line xenograft)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification prostate cancer cell line xenograft
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
Username

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投稿 Dec 17 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
Sample Mouse Cell (Neurons)
Specification Neurons
Permeabilization Yes - 0.5% Triton X100
Fixative Paraformaldehyde
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投稿 Jul 31 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Sample Mouse Tissue lysate - whole (Liver)
Specification Liver
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Jul 22 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Sample Zebrafish Tissue sections (Skin)
Specification Skin
Permeabilization No
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Jan 11 2014

1-10 of 35 Abreviews or Q&A

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