Invitrogen

PCNA Monoclonal Antibody (PC10), Biotin

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

3 Published Figures

13 References

View all (102) PCNA antibodies

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Protocols

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PCNA Antibody in Western Blot (WB) — Western blot analysis of PCNA was performed by loading 20 µg of HeLa (lane1), Raji (lane2), MCF-7 (lane3), Jurkat (lane4), HEK-293 (lane5), NIH/3T3 (lane6), MDA-MB-231 (lane7) cell lysate using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. PCNA was detected at ~ 34 kDa using PCNA, Biotin conjugate Mouse Monoclonal Antibody (Product # 13-3940) at 1:1000 dilution in 5 % skim milk at 4°C overnight on a rocking platform. Streptavidin - HRP (Product # SA1007) at 1:2000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

PCNA Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of PCNA-Biotin showing staining in the nucleus of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a PCNA-Biotin monoclonal antibody (Product # 13-3940) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

PCNA Antibody in Immunocytochemistry (ICC/IF) — Figure 1 In vitro oncogenic transformation of primary pancreatic organoids and in vivo tumorigenesis (a) In vitro culture of wild-type neonatal pancreatic organoids. Top row. Time course stereomicroscopy demonstrating progressive growth of primary pancreatic organoids at days 7, 14 and 21 of air-liquid interface culture. Scale bars, 1 mm. Bottom row. Left, day 7 H&E staining, histologic characterization demonstrates a well-differentiated cystic epithelium with surrounding fibroblastic stroma. Scale bar, 50 mum. Right middle, phase contrast. Right bottom, adeno Cre-GFP infection of pancreatic organoids. Scale bar, 1 mm. (b) Immunofluorescence demonstrates a well-differentiated cystic epithelium with surrounding fibroblastic stroma and a preponderance of epithelial E-cadherin + and Pdx1 + ductal structures with PCNA + proliferative cells and SMA + stromal cells. Bottom row, somatostatin + and insulin + endocrine cells in a rare islet structure. Day 7 immunofluorescence is depicted. Scale bar, 25 mum. (c) Transformation of primary pancreatic organoid cultures in vitro . Pancreatic organoids from neonatal mice bearing LSL Kras G12D or/and p53 flox/flox alleles (K, P, KP) were cultured with adenovirus Cre-GFP at d0 of primary plating. Left column. Stereomicroscopy of K, P and KP organoids after 20 days of primary culture and an additional 30 days of secondary passage (50 days total). Scale bars, 5 mm. Middle column. H&E staining for the indicated genotypes at day 50. As opposed to

PCNA Antibody in Immunohistochemistry (IHC) — Published figure using PCNA monoclonal antibody (Product # 13-3940) in Immunohistochemistry

PCNA Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with PCNA siRNA and loss of signal was observed in Western Blot using Anti-PCNA monoclonal Antibody (Product # 13-3940). {KD}

Product Details

13-3940

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	View 1 publication 1 publication
Immunohistochemistry (Paraffin) (IHC (P))	1:10-1:100	View 4 publications 4 publications
Immunohistochemistry (IHC)	-	View 5 publications 5 publications
Immunocytochemistry (ICC/IF)	-	View 2 publications 2 publications
Miscellaneous PubMed (Misc)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Insect, Mouse, Non-human primate, Rat, Yeast
Published species	Human, Mouse, Sheep
Host/Isotype	Mouse / IgG2a
Class	Monoclonal
Type	Antibody
Clone	PC10
Immunogen	Rat recombinant PCNA polypeptides.
Conjugate	Biotin FITC Unconjugated
Form	Liquid
Concentration	0.2 mg/mL
Purification	purified
Storage buffer	PBS, pH 7.4, with 50% glycerol, 1% BSA
Contains	0.1% sodium azide
Storage conditions	4° C
RRID	AB_2533017

Target Information

PCNA (polymerase delta auxiliary protein) is essential for DNA replication and is involved in DNA excision and mismatch repair pathways. PCNA binds to the CDK inhibitor p21, the structure-specific endonucleases Fen1 and XPG, and DNA cytosine 5-methyltransferase (MCMT). PCNA is a potentially useful marker of cells with proliferative potential and for identifying the proliferation status of tumor tissue (i.e. relevant to prognosis). PCNA is a marker for cells in early G1 phase and S phase of the cell cycle. PCNA is found in the nucleus and is a cofactor of DNA polymerase delta, and acts as a homotrimer and helps increase the processing of leading strand synthesis during DNA replication. In response to DNA damage, PCNA is ubiquitinated and is involved in the RAD6-dependent DNA repair pathway. Two transcript variants encoding the same protein have been found for PCNA. Pseudogenes of PCNA have been described on chromosome 4 and on the X chromosome.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

HMGB1 amplifies ILC2-induced type-2 inflammation and airway smooth muscle remodelling.

PLoS pathogens

Loh Z,Simpson J,Ullah A,Zhang V,Gan WJ,Lynch JP,Werder RB,Sikder AA,Lane K,Sim CB,Porrello E,Mazzone SB,Sly PD,Steptoe RJ,Spann KM,Sukkar MB,Upham JW,Phipps S

13-3940 was used in Immunohistochemistry to investigate the processes that mediate tissue damage-induced type-2 inflammation and the origins of airway remodelling, using a preclinical mouse model of viral bronchiolitis.

Wed Jul 01 00:00:00 EDT 2020

The cJUN NH₂-terminal kinase (JNK) signaling pathway promotes genome stability and prevents tumor initiation.

eLife

Girnius N,Edwards YJ,Garlick DS,Davis RJ

13-3940 was used in Immunohistochemistry to identify cJUN NH2-terminal kinase pathway defects as 'driver' mutations that promote genome instability and tumor initiation.

Fri Jun 01 00:00:00 EDT 2018

JNK Promotes Epithelial Cell Anoikis by Transcriptional and Post-translational Regulation of BH3-Only Proteins.

Cell reports

Girnius N,Davis RJ

13-3940 was used in Immunohistochemistry-immunofluorescence to study the role of the cJUN NH2-terminal kinase signalling pathway in anoikis.

Tue Nov 14 00:00:00 EST 2017

Oncogenic transformation of diverse gastrointestinal tissues in primary organoid culture.

Nature medicine

Li X,Nadauld L,Ootani A,Corney DC,Pai RK,Gevaert O,Cantrell MA,Rack PG,Neal JT,Chan CW,Yeung T,Gong X,Yuan J,Wilhelmy J,Robine S,Attardi LD,Plevritis SK,Hung KE,Chen CZ,Ji HP,Kuo CJ

13-3940 was used in immunohistochemistry - paraffin section to study the oncogenic transformation in primary organoid culture of gastrointestinal tissues

Tue Jul 01 00:00:00 EDT 2014

Oncogenic transformation of diverse gastrointestinal tissues in primary organoid culture.

Nature medicine

Li X,Nadauld L,Ootani A,Corney DC,Pai RK,Gevaert O,Cantrell MA,Rack PG,Neal JT,Chan CW,Yeung T,Gong X,Yuan J,Wilhelmy J,Robine S,Attardi LD,Plevritis SK,Hung KE,Chen CZ,Ji HP,Kuo CJ

13-3940 was used in immunohistochemistry - paraffin section to study the oncogenic transformation in primary organoid culture of gastrointestinal tissues