Anti-PCNA antibody [EPR3821] Recombinant (ab92552)

Product name

Anti-PCNA antibody [EPR3821]
See all PCNA primary antibodies
Description

Rabbit monoclonal [EPR3821] to PCNA
Host species

Rabbit
Tested applications

Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide within Human PCNA aa 100-200. The exact sequence is proprietary.
Database link: P12004
Positive control
- HeLa, HepG2, 293T and A431 cell lysates, Tonsil tissue, HeLa cells
General notes

A trial size is available to purchase for this antibody.

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR3821
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Positive Controls
- HeLa nuclear extract lysate (ab14655)
- HepG2 nuclear extract lysate (ab14660)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

Our Abpromise guarantee covers the use of ab92552 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		1/1000 - 1/10000. Predicted molecular weight: 29 kDa.
IP		1/20 - 1/50.
IHC-P		1/100 - 1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. See IHC antigen retrieval protocols. The use of an HRP/AP polymerized antibody is recommended for a secondary antibody.
Flow Cyt		1/40. For unpurified, use 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF		1/100 - 1/250. Use with methanol fixed samples.

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 accession P12004 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 5111 Human
- Entrez Gene: 18538 Mouse
- Entrez Gene: 25737 Rat
- Omim: 176740 Human
- SwissProt: P12004 Human
- SwissProt: P17918 Mouse
- SwissProt: P04961 Rat
- Unigene: 147433 Human
- Unigene: 728886 Human
- Unigene: 7141 Mouse
- Unigene: 223 Rat
see all
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

Western blot - Anti-PCNA antibody [EPR3821] (ab92552)

All lanes : Anti-PCNA antibody [EPR3821] (ab92552) at 1/1000 dilution (purified)

Lane 1 : mouse spleen lysate
Lane 2 : NIH/3T3 lysate
Lane 3 : PC-12 lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

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

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)

Immunohistochemical staining of paraffin embedded rat liver with purified ab92552 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)

Immunohistochemical staining of paraffin embedded mouse testis with purified ab92552 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)

Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab92552 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)

Unpurified ab92552 showing positive staining in Normal colon tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)

Unpurified ab92552 showing positive staining in Urinary bladder carcinoma tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)

Unpurified ab92552 showing positive staining in Ovarian carcinoma tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)

Unpurified ab92552 showing positive staining in Breast carcinoma tissue.
Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody [EPR3821] (ab92552)Image courtesy of an abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada

Unpurified ab92552 (1/200) staining PCNA in Hela cells (green). Cells were fixed in methanol and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to abreview.
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody [EPR3821] (ab92552)

Immunofluorescence staining of A431 cells with purified ab92552 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab92552 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody [EPR3821] (ab92552)

Unpurified ab92552 at 1/100 dilution staining PCNA in HeLa cells by Immunofluorescence.
Western blot - Anti-PCNA antibody [EPR3821] (ab92552)

All lanes : Anti-PCNA antibody [EPR3821] (ab92552) at 1/5000 dilution (purified)

Lane 1 : HeLa cell lysate
Lane 2 : HEK293 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

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

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-PCNA antibody [EPR3821] (ab92552)

All lanes : Anti-PCNA antibody [EPR3821] (ab92552) at 1/50000 dilution (unpurified)

Lane 1 : HeLa cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : 293T cell lysate
Lane 4 : A431 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 29 kDa
Immunoprecipitation - Anti-PCNA antibody [EPR3821] (ab92552)

ab92552 (purified) at 1/20 immunoprecipitating PCNA in 10 μg HeLa (Lanes 1 and 2, observed at 29 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
Flow Cytometry - Anti-PCNA antibody [EPR3821] (ab92552)

Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified ab92552 at a dilution of 1 in 40 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
Flow Cytometry - Anti-PCNA antibody [EPR3821] (ab92552)

Overlay histogram showing HeLa cells stained with unpurified ab92552 (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 (ab92552, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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.

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Liu W et al. Dioscin inhibits stem-cell-like properties and tumor growth of osteosarcoma through Akt/GSK3/ß-catenin signaling pathway. Cell Death Dis 9:343 (2018). Read more (PubMed: 29497056) »
Yang Z et al. Anticancer effect of fufang yiliu yin on human hepatocellular carcinoma SMMC-7721 cells. Am J Transl Res 10:491-500 (2018). Read more (PubMed: 29511443) »

See all 32 Publications for this product

Publishing research using ab92552? Please let us know so that we can cite the reference in this datasheet.

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Anti-PCNA antibody [EPR3821] (ab92552)

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Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PCNA antibody [EPR3821]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PCNA antibody [EPR3821]

Immunocytochemistry/ Immunofluorescence abreview for Anti-PCNA antibody [EPR3821]