Recombinant Anti-PCNA antibody [EPR3821] (ab92552)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3821] to PCNA
- Suitable for: WB, IP, Flow Cyt (Intra), IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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, Flow Cyt (Intra), IHC-P, 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
- WB: Mouse spleen lysate. HeLa, NIH/3T3, PC-12, HepG2, HEK-293, HEK-293T and A431 cell lysates. IHC-P: Human ovarian carcinoma, urinary bladder carcinoma, normal colon, breast carcinoma and cervical carcinoma tissue. Rat liver tissue. Mouse testis tissue. ICC/IF: A431 and HeLa cells. IP: HeLa cell lysate. Flow Cyt (intra): HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3821 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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WB | (5) |
1/1000 - 1/10000. Predicted molecular weight: 29 kDa.
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IP |
1/20 - 1/50.
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Flow Cyt (Intra) |
1/40.
For unpurified, use 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P | (4) |
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. |
ICC/IF | (1) |
1/100 - 1/250.
Use with methanol fixed samples. |
Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 29 kDa. |
IP
1/20 - 1/50. |
Flow Cyt (Intra)
1/40. For unpurified, use 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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. |
ICC/IF
1/100 - 1/250. Use with methanol fixed samples. |
Target
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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
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 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
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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
see all -
Alternative names
- ATLD2 antibody
- cb16 antibody
- Cyclin antibody
see all
Images
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All lanes : Anti-PCNA antibody [EPR3821] (ab92552) at 1/1000 dilution (purified)
Lane 1 : Mouse spleen lysate
Lane 2 : NIH/3T3 (Mouse embryo fibroblast cell line) lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) 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 kDaBlocking 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.
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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.
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)
Unpurified ab92552 showing positive staining in human normal colon tissue.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)
Unpurified ab92552 showing positive staining in human urinary bladder carcinoma tissue.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)
Unpurified ab92552 showing positive staining in human ovarian carcinoma tissue.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [EPR3821] (ab92552)
Unpurified ab92552 showing positive staining in human breast carcinoma tissue.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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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.
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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.
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Unpurified ab92552 at 1/100 dilution staining PCNA in HeLa cells by Immunofluorescence.
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All lanes : Anti-PCNA antibody [EPR3821] (ab92552) at 1/5000 dilution (purified)
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) 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 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
All lanes : Anti-PCNA antibody [EPR3821] (ab92552) at 1/50000 dilution (unpurified)
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
Lane 3 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lane 4 : A431 (Human epidermoid carcinoma cell line) 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 -
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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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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).
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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 Fluorr® 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (354)
ab92552 has been referenced in 354 publications.
- Li J & Chen H Actin-binding Rho activating C-terminal like (ABRACL) transcriptionally regulated by MYB proto-oncogene like 2 (MYBL2) promotes the proliferation, invasion, migration and epithelial-mesenchymal transition of breast cancer cells. Bioengineered 13:9019-9031 (2022). WB ; Human . PubMed: 35341461
- Xu J et al. Uncarboxylated osteocalcin promotes proliferation and metastasis of MDA-MB-231 cells through TGF-ß/SMAD3 signaling pathway. BMC Mol Cell Biol 23:18 (2022). WB ; Human . PubMed: 35413833
- Liu Y et al. CircRIP2 promotes NSCLC progression by sponging for miR-671-5p to regulate FOXM1 expression. Histol Histopathol 37:117-124 (2022). PubMed: 34291441
- Xu D et al. miR-143-3p represses leukemia cell proliferation by inhibiting KAT6A expression. Anticancer Drugs 33:e662-e669 (2022). PubMed: 34459452
- Li F et al. CircAKT3 promotes cell proliferation, survival and glutamine metabolism of gastric cancer by activating SLC1A5 expression via targeting miR-515-5p. Histol Histopathol 37:227-241 (2022). PubMed: 34850965