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Products:Cell Biology >> Cell Cycle >> Markers
Overview
Product name
Anti-PCNA antibody - Proliferation Marker
PCNA 抗体 (29件) 一覧
Description
Rabbit polyclonal to PCNA - Proliferation Marker
Tested applications
WB, ICC/IF, IHC-P, IHC-FoFr, IPmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Predicted to work with
Sheep, Cow, Dog, Xenopus laevis
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 200 to the C-terminus of Human PCNA.
(Peptide available as ab18602.)
Positive control
This antibody gave a positive control in the following human whole cell lysate: HEK 293 (Human embryonic kidney cell line) This antibody gave a positive control in the following mouse whole cell lysates: NIH 3T3 (Mouse embryonic fibroblast cell line) MEF1 (Mouse embryonic fibroblast cell line)
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: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
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
Applications
Show applications key
Our Abpromise guarantee covers the use of ab18197 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).Can be blocked with PCNA peptide (ab18602).
ICC/IF: Use a concentration of 1 µg/ml
IHC-P: Use at an assay dependent concentration.
IHC-FoFr: Use at an assay dependent concentration. (PubMed: 21895533)
IP: Use at an assay dependent concentration.
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).
Alternative names
- Cyclin antibody
- Cyclin antibody
- DNA polymerase delta auxiliary protein antibody
see all
Database links
- Entrez Gene: 5111 Human
- Entrez Gene: 18538 Mouse
- Entrez Gene: 25737 Rat
- Entrez Gene: 398425 Xenopus laevis
- IMAGE Id: 2820036 Human
- Omim: 176740 Human
- SwissProt: P12004 Human
- SwissProt: P17918 Mouse
see all
Anti-PCNA antibody - Proliferation Marker images:
Western blot
All lanes : Anti-PCNA antibody - Proliferation Marker (ab18197) at 1 µg/ml
Lane 1 : HEK293 lysate
Lane 2 : HEK293 lysate with PCNA peptide (ab18602) at 1 µg/ml
Lysates/proteins at 20 µg/ml per lane.
Secondary
Alexa Fluor Goat-anti-Rabbit IgG (700) at 1/10000 dilution
Predicted band size : 29 kDa
Observed band size : 29 kDa
Additional bands at : 48 kDa (possible cross reactivity),75 kDa (possible cross reactivity).
Immunocytochemistry/ Immunofluorescence - PCNA antibody (ab18197)
ICC/IF image of ab18197 stained NIH/3T3 cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18197, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Panel A does not show the Alexa Fluor® 488 channel, Panel B shows the specfic nuclear staining by ab18197.Immunocytochemistry/ Immunofluorescence - PCNA antibody (ab18197)
ab18197, at a 1/5000 dilution, staining PCNA in assynchronous HeLa cells. Cells were counter-stained with DAPI (red). For more information please refer to Abreview.
This image is courtesy of an Abreview submitted by Dr Kirk McManus
Western blot - PCNA antibody (ab18197)
All lanes : Anti-PCNA antibody - Proliferation Marker (ab18197) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 29 kDa
Observed band size : 29 kDa
Additional bands at : 48 kDa (possible cross reactivity),52 kDa. We are unsure as to the identity of these extra bands.
Immunocytochemistry/ Immunofluorescence - PCNA antibody (ab18197)
ab18197, at a 1/2000 dilution staining PCNA in MRC5 Sv40 transformed fibroblasts. Cells were counterstained with DAPI (blue). For more information please refer to Abreview.
Dr Catherine Green
Immunoprecipitation - PCNA antibody (ab18197)
ab18197 staining rat PC12 whole cell lysate by Immunoprecipitation.
ab18197 was incubated with the lysate (at a concentration of 5µg/ml) and a Protein A matrix for 12 hours at 4°C to achieve immunoprecipitation. 400µg of protein were present in the lysate input.
Lane order: PCNA IP (lane 1), Control IP (lane 2), Input 5% (lane 3)
This antibody immunoprecipitates a product of ~34 kDa which is consistent with the mobility of PCNA on SDS-PAGE
ab18197 was also used for the western blot step, at a contration of 1µg/ml
This image is courtesy of an Abreview submitted by Antibody Solutions Ltd
Immunocytochemistry/ Immunofluorescence - PCNA antibody - Proliferation Marker (ab18197)
ICC/IF image of ab18197 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 (ab18197, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) 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.
Western blot
Anti-PCNA antibody - Proliferation Marker (ab18197) at 1 µg/ml + PCNA protein (His tag) (ab85651) at 0.01 µg
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 10 seconds
References for Anti-PCNA antibody - Proliferation Marker (ab18197)
This product has been referenced in:
- Kabadi SVet al. Cyclin D1 Gene Ablation Confers Neuroprotection in Traumatic Brain Injury. J Neurotrauma : (2011). IHC-FoFr; Mouse.Read more (PubMed: 21895533) »
- Eo Jet al. Complex ovarian defects lead to infertility in Etv5-/- female mice. Mol Hum Reprod 17:568-76 (2011).Read more (PubMed: 21478265) »
See all 9 publications for this product
Publishing research using ab18197? 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"