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AB264494

Anti-PCNA antibody [PC10] - BSA and Azide free

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(1 Publication)

Mouse Recombinant Monoclonal PCNA antibody. Carrier free. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Rat, Mouse, Human samples. Cited in 1 publication.

View Alternative Names

DNA sliding clamp PCNA, Cyclin, Proliferating cell nuclear antigen, PCNA

5 Images
Flow Cytometry (Intracellular) - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)
  • Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)

Flow cytometry overlay histogram showing HeLa cells stained with ab264494 (red line). The cells were fixed with 4 % formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab264494) (1x106 in 100 μL at 1 μg/ml) for 30 min at 22°C.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 22°C.

Isotype control antibody (black line) was mouse IgG2aκ; (ab18413) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)

ab29 stained in Hela cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab29 at 5 μg/ml and ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150117 (pseudo-colored red) and ab150080 (colored green) used at 1 μg/ml for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 μM for 1hour at room temperature.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab29).

Flow Cytometry (Intracellular) - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)

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 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. 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.

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 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113)at 1/2000 dilution for 30 min at 22°C. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab29).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)
  • IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)

IHC image of PCNA staining in a section of formalin-fixed paraffin-embedded normal rat large intestine 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 20mins. The section was then incubated with ab264494, 0.1ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)
  • IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCNA antibody [PC10] - BSA and Azide free (AB264494)

IHC image of PCNA staining in a section of formalin-fixed paraffin-embedded normal spleen 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 20mins. The section was then incubated with ab264494, 0.1ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

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.

Key facts

Host species

Mouse

Clonality

Monoclonal

Clone number

PC10

Isotype

IgG2a

Light chain type

kappa

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

ICC/IF, Flow Cyt (Intra), IHC-P, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IHCP-species-checked": "guaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p>Methanol fixation recommended</p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/mouse-igg2c-kappa-monoclonal-18c8bc7ad10-isotype-control-ab170191'>ab170191</a> - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.</p>" }, "Mouse": { "IHCP-species-checked": "guaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/mouse-igg2c-kappa-monoclonal-18c8bc7ad10-isotype-control-ab170191'>ab170191</a> - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.</p>" }, "Rat": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p>Methanol fixation recommended</p>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/mouse-igg2c-kappa-monoclonal-18c8bc7ad10-isotype-control-ab170191'>ab170191</a> - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.</p>" }, "Catshark": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Chicken": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Cow": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Dogfish": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Drosophila melanogaster": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Monkey": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Pig": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Pigeon": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Thornback ray": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Zebrafish": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" } } }
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Product details

ab264494 is the carrier-free version of ab29.

Want a custom formulation?
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purity
IgG fraction
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

PCNA or Proliferating Cell Nuclear Antigen functions as a sliding clamp for DNA polymerases during DNA replication. This important role enables it to coordinate the orderly duplication of the genome by increasing the processivity of DNA polymerase δ. PCNA weighs approximately 29 kDa and forms a homotrimeric ring structure. It is expressed in the nucleus of cells often found abundantly in proliferating cells. Aside from its main name some people also refer to it as PC10.
Biological function summary

This protein serves a core function in cell cycle regulation and DNA repair. PCNA acts as a scaffold for the assembly of numerous proteins involved in DNA processing forming part of the replication fork complex. It interacts with cyclins and cyclin-dependent kinases (CDKs) linking DNA synthesis to cell cycle progression. Furthermore PCNA partners with proteins involved in DNA mismatch repair base excision repair and nucleotide excision repair.

Pathways

PCNA is integral in the DNA replication and repair pathways. It closely associates with the p21 protein which regulates the cell cycle by inhibiting cyclin-CDK complexes upon DNA damage. PCNA also plays a part in the ubiquitin-proteasome pathway where its modification by ubiquitin impacts how cells respond to DNA damage. This modification recruits specific proteins for DNA repair highlighting its central role in maintaining genomic stability.

PCNA is linked to cancer and autoimmune diseases. Its overexpression is common in various cancers reflecting its role in cell proliferation and the cell cycle. PCNA antibodies are sometimes found in the serum of patients with systemic lupus erythematosus (SLE) a connection also related to alterations in the p21 protein pathway. The interaction between PCNA and disease-specific proteins emphasizes its importance in both cell proliferation and immune system disorders.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Confers DNA tethering and processivity to DNA polymerases and other proteins (PubMed : 24695737, PubMed : 24939902, PubMed : 35585232). Auxiliary protein of DNA polymerase delta and epsilon, is involved in the control of DNA replication by increasing the polymerases' processivity during elongation of the leading strand (PubMed : 35585232). 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. Plays a key role in DNA damage response (DDR) by being conveniently positioned at the replication fork to coordinate DNA replication with DNA repair and DNA damage tolerance pathways (PubMed : 24939902). Acts as a loading platform to recruit DDR proteins that allow completion of DNA replication after DNA damage and promote postreplication repair : monoubiquitinated PCNA leads to recruitment of translesion (TLS) polymerases, while 'Lys-63'-linked polyubiquitination of PCNA is involved in error-free pathway and employs recombination mechanisms to synthesize across the lesion (PubMed : 24695737).
See full target information PCNA
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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Cell reports 38:110220 PubMed35021081

2022

2-Hydroxyglutarate destabilizes chromatin regulatory landscape and lineage fidelity to promote cellular heterogeneity.

Applications

Unspecified application

Species

Unspecified reactive species

Meena Kusi,Maryam Zand,Li-Ling Lin,Meizhen Chen,Anthony Lopez,Chun-Lin Lin,Chiou-Miin Wang,Nicholas D Lucio,Nameer B Kirma,Jianhua Ruan,Tim H-M Huang,Kohzoh Mitsuya
View all publications
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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