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Anti-PCNA antibody [PC10] - Proliferation Marker (ab29)

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Overview

Product name

Anti-PCNA antibody [PC10] - Proliferation Marker
See all PCNA products (29) ...

Description

Mouse monoclonal [PC10] to PCNA - Proliferation Marker

Specificity

This antibody is specific for PCNA p36 protein expressed at high levels in proliferating cells.

Tested applications

IHC-P, IP, WB, ICC/IF, IHC-Fr, Flow Cytmore details

Cross reactivity

Reacts with

Mouse, Rat, Chicken, Human, Fruit fly (Drosophila melanogaster), Zebrafish

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

Properties

Form

Liquid

Storage instructions

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: 0.02% Sodium Azide
Constituents: PBS

Concentration

Concentration information loading...

Purity

IgG fraction

Clonality

Monoclonal

Clone number

PC10

Myeloma

Sp2/0-Ag14

Isotype

IgG2a

Light chain type

kappa

Research areas

Cancer >> Tumor biomarkers >> Tumor antigens
Cancer >> Cell cycle >> Cell division
Epigenetics and Nuclear Signaling >> Chromatin Binding Proteins >> DNA / RNA binding
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Synthesis >> Other
Tags & Cell Markers >> Cell Type Markers >> Replication
Epigenetics and Nuclear Signaling >> DNA methylation >> Methylated DNA Binding
Cell Biology >> Cell Cycle >> Markers

  • Immunohistochemistry (Frozen sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)Immunohistochemistry (Frozen sections) - PCNA antibody [PC10] - Proliferation Marker (ab29) image (enlarge)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29) image (enlarge)

  • Western blot - PCNA antibody [PC10] - Proliferation Marker (ab29)Western blot - PCNA antibody [PC10] - Proliferation Marker (ab29) image (enlarge)

Applications

Show applications key

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.

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.

Target information above from: UniProt accessionP12004 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

Alternative names

  • Cyclin antibody
  • DNA polymerase delta auxiliary protein antibody
  • HGCN8729 antibody
  • MGC8367 antibody
  • Mutagen-sensitive 209 protein antibody
  • PCNA antibody
  • Pcna/cyclin antibody
  • PCNA_HUMAN antibody
  • PCNAR antibody
  • Polymerase delta accessory protein antibody
  • Proliferating Cell Nuclear Antigen antibody
see all

Database links

see all

Anti-PCNA antibody [PC10] - Proliferation Marker images:

  Immunohistochemistry (Frozen sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunohistochemistry (Frozen sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

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.

Carl Hobbs, CARD, KCL, UK

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

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).

Carl Hobbs, CARD, KCL, UK

  Western blot - PCNA antibody [PC10] - Proliferation Marker (ab29)

Western blot - PCNA antibody [PC10] - Proliferation Marker (ab29)

All lanes : Anti-PCNA antibody [PC10] - Proliferation Marker (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

  Flow Cytometry - Anti-PCNA antibody [PC10] - Proliferation Marker (ab29)

Flow Cytometry - Anti-PCNA antibody [PC10] - Proliferation Marker (ab29)

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 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, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  Western blot - PCNA antibody [PC10] - Proliferation Marker (ab29)

Western blot - PCNA antibody [PC10] - Proliferation Marker (ab29)

All lanes : Anti-PCNA antibody [PC10] - Proliferation Marker (ab29) at 1/5000 dilution

Lane 1 : Human 293 total cell extract
Lane 2 : Chicken DT40 total cell extract


Performed under reducing conditions.

Predicted band size : 29 kDa
Observed band size : 30 kDa (why is the actual band size different from the predicted?)


Exposure time : 5 seconds

Blocking for 2 hours in 5% milk. Primary antibody incubated for 1 hour.

Taken from an Abreview submitted by Javier di Noia

  Immunoprecipitation - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunoprecipitation - PCNA antibody [PC10] - Proliferation Marker (ab29)

ab29 at a 1/500 dilution used in immunoprecipitation.

Lane 1: Beads only + HeLa whole cell lysate

Lane 2: Beads only + SupT1 whole cell lysate

Lane 3: Beads + ab29 + HeLa whole cell lysate

Lane 4: Beads + ab29 + SupT1 whole cell lysate

A 50KDa band is precipitated which is IgG heavy chain whilst the 30kDa band is PCNA.

This image is courtesy of an anonymous Abreview

See Abreview

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

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.

This image is courtesy of an Abreview submitted by Mr Carl hobbs

See Abreview

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

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.

Carl Hobbs, CARD, KCL, UK

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

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).

Carl Hobbs, CARD, KCL, London, UK

  Immunocytochemistry/ Immunofluorescence - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunocytochemistry/ Immunofluorescence - PCNA antibody [PC10] - Proliferation Marker (ab29)

ICC/IF image of ab29 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab29, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HEK 293, HepG2 and MCF7 cells.

  Immunohistochemistry (Frozen sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunohistochemistry (Frozen sections) - PCNA antibody [PC10] - Proliferation Marker (ab29)

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.

This image is a courtesy of Anonymous Abreview

See Abreview

  Immunocytochemistry/ Immunofluorescence - PCNA antibody [PC10] - Proliferation Marker (ab29)

Immunocytochemistry/ Immunofluorescence - PCNA antibody [PC10] - Proliferation Marker (ab29)

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 DyLight® 488 goat anti-mouse 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.

References for Anti-PCNA antibody [PC10] - Proliferation Marker (ab29)

This product has been referenced in:

  • Soleimani Ret al. Enhancement of neoangiogenesis and follicle survival by sphingosine-1-phosphate in human ovarian tissue xenotransplants. PLoS One 6:e19475 (2011). IHC-P; Human.Read more (PubMed: 21559342) »
  • Beck Het al. Regulators of cyclin-dependent kinases are crucial for maintaining genome integrity in S phase. J Cell Biol 188:629-38 (2010). WB; Human.Read more (PubMed: 20194642) »

See all 39 publications for this product

Publishing research using ab29? Please let us know so that we can cite the reference in this datasheet

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"