Overview

  • Product name
    Anti-PCNA antibody [PC10]
    See all PCNA primary antibodies
  • Description
    Mouse monoclonal [PC10] to PCNA
  • Specificity
    This antibody is specific for PCNA p36 protein expressed at high levels in proliferating cells.
  • Tested applications
    Suitable for: IHC-P, IP, WB, ICC/IF, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Pigeon, Human, Pig, Drosophila melanogaster, Monkey, Zebrafish, Thornback ray, Dogfish, Catshark
    Predicted to work with: Cow
  • Immunogen

    Protein A-rat PCNA (proliferating cell nuclear antigen) fusion obtained from pC2T

  • Positive control
    • WB: DT40 B lymphoma cell lysate, 293 cell lysate (see review), Hela, HEK293, A431 whole cell lysates. IHC-P: mouse hippocampus (see review), Normal human tonsil, developing chick brain. IHC-Fr: rat intestine. Flow Cyt: HeLa.
  • General notes

    This antibody clone [PC10] is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab29 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
IHC-P 1/10000 - 1/30000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 29 kDa).
ICC/IF Use a concentration of 0.5 - 1 µg/ml. Methanol fixation recommended
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the PCNA family.
  • Post-translational
    modifications
    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.
  • Cellular localization
    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
  • Database links
  • Alternative names
    • 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

Images

  • All lanes : Anti-PCNA antibody [PC10] (ab29) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 29 kDa
    Observed band size : 29 kDa


    Exposure time : 4 minutes
  • ab29 staining PCNA in mouse embryonic brain tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with paraformaldehyde and permeabilized with 0.1% PBS-Tween before blocking with 5% BSA for 1 hour at 220C. The sample was incubated with primary antibody (1/500) in 5% BSA in 0.3% PBS-Triton-X100 for 14 hours at 220C. An Alexa Fluor®488-conjugated Goat polyclonal to mouse IgG was used as secondary antibody at 1/500 dilution.

    See Abreview

  • ICC/IF image of ab29 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 (ab29, 5µg/ml) overnight at +4°C. The secondary antibody (green) was goat anti-mouse DyLight® 488 (IgG - H&L, pre-adsorbed) (ab96879) 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.

  • Overlay histogram showing HeLa cells stained with ab29 (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 (ab29, 0.1μg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 0.1μg/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 image of PCNA staining in rat large intestine formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab29, 0.025µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • IHC image of PCNA staining in rat spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab29, 1/30,000 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Mouse monoclonal [PC10] to PCNA - Proliferation Marker (ab29) used in immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections; 1/6000 for 2h at RT) on Human Tissue sections (Paget's disease of the nipple). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Incubation time: Secondary Antibody: Biotin conjugated goat anti mouse Igs (1/200).
  • ab29 at 1/6000 staining mouse embryo (day 17) liver and gut tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in Tris buffer was performed. The tissue was blocked before incubation with the antibody for 2 hours. A biotinylated goat polyclonal antibody was used as the secondary.

    See Abreview

  • Mouse monoclonal [PC10] to PCNA - Proliferation Marker (ab29) used in immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections; 1/6000 for 2h at RT) on E6 developing chick brain). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Incubation time: Secondary Antibody: Biotin conjugated goat anti mouse Igs (1/200). NB: This image shows developing brain/overlying skin.
  • Immunohistochemistical staining (Formaldehyde/PFA-fixed paraffin-embedded sections) for PCNA antibody [PC10] - Proliferation Marker (ab29) on Rat Tissue sections (adult spinal cord DRG). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Primary Antibody used at 1/6000 for 2 minutes at RT. Secondary Antibody: Biotin labelled goat anti mouse Igs (1/200).
  • Mouse monoclonal [PC10] to PCNA - Proliferation Marker (ab29) used in immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections;1/6000 for 2h at RT) on intestine of adult Zebra fish). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Incubation time: Secondary Antibody: Biotin conjugated goat anti mouse Igs (1/200). NB: The crypt nuclei on this image of zebrafish intestine, are positive for the PCNA/PC10 clone conforming to accepted localisation data for PCNA in other species.

References

This product has been referenced in:
  • Yu J  et al. REC8 functions as a tumor suppressor and is epigenetically downregulated in gastric cancer, especially in EBV-positive subtype. Oncogene 36:182-193 (2017). WB ; Human . Read more (PubMed: 27212034) »
  • Sun N  et al. Inhibitory effect of dexamethasone on residual Lewis lung cancer cells in mice following palliative surgery. Oncol Lett 13:356-362 (2017). WB, IHC-P ; Mouse . Read more (PubMed: 28123567) »

See all 200 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Carp Tissue sections (Testis)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Permeabilization
No
Specification
Testis
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde
Username

Emi Tucker

Verified customer

Submitted Oct 19 2017

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Nothobranchius furzeri Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Jolien Van houcke

Verified customer

Submitted Apr 05 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (EC109)
Gel Running Conditions
Non-reduced Denaturing (10%SDS)
Loading amount
80 µg
Specification
EC109
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Qingyuan Yang

Verified customer

Submitted Jan 08 2016

Application
Western blot
Sample
Rat Tissue lysate - whole (Brain homogenate)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
Brain homogenate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 20 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Fish Tissue sections (skeletal muscle of pseudoplatystoma fasciatum)
Antigen retrieval step
None
Permeabilization
No
Specification
skeletal muscle of pseudoplatystoma fasciatum
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Acetone
Username

Dr. Danilo Aguiar

Verified customer

Submitted May 27 2015



The immunogen used to produce ab29 is full length rat PCNA fused to Protein A. The exact epitope recognized by ab29 has not been determined.

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Dako Protein Block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
Sample
Pig Cell (Intestinal epithelium)
Specification
Intestinal epithelium
Permeabilization
Yes - 0.3% PBS-T
Fixative
Paraformaldehyde
Username

Dr. Liara Gonzalez

Verified customer

Submitted Feb 05 2014

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - whole cell (HCT116)
Specification
HCT116
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 19 2013

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 25°C
Sample
Rat Cell (Oliogdendorcytes)
Specification
Oliogdendorcytes
Permeabilization
Yes - 0.01% Triton X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 10 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
2% normal goat serum, 1% bovine serum albumin, 0.1% cold fish skin gelatine, 0.1% Triton X-100, 0.05% Tween 20, 0.05% sodium azide, 0.01M PBS, pH 7.2 as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer (pH 9.0)
Sample
Frog Tissue sections (Proximal Small intestine)
Specification
Proximal Small intestine
Permeabilization
No
Fixative
Formaldehyde
Username

Dr. Rebecca Cramp

Verified customer

Submitted Nov 22 2013

1-10 of 86 Abreviews or Q&A

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