Product datasheet

Anti-PCNA antibody [PC10] ab29
72 Abreviews 736 References 画像数 14
製品の概要
製品名

Anti-PCNA antibody [PC10]

製品の詳細

Mouse monoclonal [PC10] to PCNA

由来種Mouse

アプリケーション

適用あり : IHC-P, WB, ICC/IF, Flow Cyt (Intra)

種交差性

交差種 : Mouse, Rat, Human
交差が予測される動物種 : Chicken, Cow, Pigeon, Pig, Drosophila melanogaster, Monkey,Zebrafish, Thornback ray, Dogfish, Catshark
免疫原

Fusion protein corresponding to PCNA. Protein A-PCNA fusion protein obtained from pC2Tconstruct. This construct lacked 93 nucleotides at the 3' end of PCNA.

ポジティブ･コントロール

WB: HeLa, HEK293, A431 whole cell lysates, PC12, SV40LT-SMC, NIH 3T3, rat liver, rat heart.
IHC-P: Human breast (Paget's disease of the nipple), mouse liver, mouse gut, rat spinal cord
DRG, developing chick brain, zebrafish intestine, rat spleen, rat large intestine. IHC-Fr: mouseembryonic brain, HeLa. Flow Cyt: HeLa.

特記事項

This antibody clone [PC10] is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for yourexperiments, please contact orders@abcam.com or you can find further information here.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years.
Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies
and knockout edited cell lines for gold-standard validation. Please check that this product meets
your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/orcontact our Support team ahead of purchase. Recommended alternatives for this product can befound below, along with publications, customer reviews and Q&As

製品の特性
製品の状態

Liquid

保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.

バッファー

pH: 7.2
Preservative: 0.02% Sodium azide
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Constituents: PBS, 6.97% L-Arginine
精製度

Protein G purified

ポリ /モノモノクローナル

クローン名PC10

ミエローマ

Sp2/0-Ag14

アイソタイプ

IgG2a

軽鎖の種類

kappa

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab29の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション

特記事項

Abreviews

IHC-P(28)

1/10000 - 1/30000. Perform heat mediated antigen retrieval withcitrate buffer pH 6 before commencing with IHC staining protocol.

WB

(20)

Use a concentration of 1 µg/ml. Detects a band of approximately
30 kDa (predicted molecular weight: 29 kDa).

ICC/IF(14)

Use a concentration of 1 - 5 µg/ml.
Methanol fixation recommended
Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an
isotype control with this antibody.

Flow Cyt (Intra)

ターゲット情報機能This protein is an auxiliary protein of DNA polymerase delta and is involved in the control ofeukaryotic DNA replication by increasing the polymerase's processibility during elongation of theleading strand. Induces a robust stimulatory effect on the 3'-5' exonuclease and 3'phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. Has tobe loaded onto DNA in order to be able to stimulate APEX2.

配列類似性

Belongs to the PCNA family.

翻訳後修飾

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 inundamaged 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 andpromotes degradation.

細胞内局在Nucleus. Forms nuclear foci representing sites of ongoing DNA replication and vary inmorphology and number during S phase. Together with APEX2, is redistributed in discrete2

nuclear foci in presence of oxidative DNA damaging agents.

画像

Immunohistochemistry analysis (Formalin/PFA-fixed paraffinembedded sections) of paraformaldehyde-fixed mouse braintissue. Stained with ab29 at 1/10000 dilution. Secondary antibody
used was ab150109 at 1/250 dilution. Blocking was done with 5%
serum for 1 hour at 21°C. The sample was incubated with theprimary antibody and 5% Normal Donkey Serum for 19 hours at4°C. Antigen retrieval method was heat mediated, Tris/EDTA.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-PCNA antibody [PC10](ab29)
This image is courtesy of an anonymous Abreview

All lanes : Anti-PCNA antibody [PC10] (ab29) at 1 µg/ml
Lane 1 : HeLa 20 ug
Lane 2 : PC12 20 ug
Lane 3 : SV40LT-SMC 20 ug
Lane 4 : NIH 3T3 20 ug
Lane 5 : Rat liver 20 ug
Lane 6 : Rat heart 20 ug
SecondaryAll lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW)
Western blot - Anti-PCNA antibody [PC10] (ab29)

preadsorbed (ab216772) at 1/20000 dilution
Performed under reducing conditions.

Predicted band size: 29 kDa

Merged signal (red and green). Green - ab29 observed at 32 kDa.
Red - loading control, ab52866, observed at 50 kDa.
All samples were subjected to SDS-PAGE. The membrane wasblocked with 3% NF Milk. Ab29 and ab52866(Rabbit anti alpha3

Tubulin loading control) were incubated overnight at 4°C at 1 ug/mL
and 1/20000 dilution respectively. Blots were developed with Goatanti-Mouse IgG H&L (IRDye® 800CW)
preabsorbed ab216772 and Goat anti-Rabbit IgG H&L(IRDye® 680RD) preabsorbed ab216777 secondary antibodies at1/20000 dilution for 1 hour at room temperature before imaging.

Mouse monoclonal [PC10] to PCNA - Proliferation Marker (ab29)
used in immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections; 1/6000 for 2h at RT) on human tissue sections(Paget's disease of the nipple). Antigen retrieval step: Heat
mediated. Blocking step: 1% BSA for 10 mins at RT. Incubationtime: Secondary Antibody: Biotin conjugated goat anti mouse Igs(1/200).

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-PCNA antibody [PC10](ab29)
This image is courtesy of Carl Hobbs, King's CollegeLondon, United Kingdom

All lanes : Anti-PCNA antibody [PC10] (ab29) at 5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell
LysateLane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell
LysateLane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell
LysateLysates/proteins at 10 µg per lane.

Western blot - Anti-PCNA antibody [PC10] (ab29)

SecondaryAll lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed(HRP) at 1/3000 dilution
Performed under reducing conditions.

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

4Exposure time: 4 minutes
ab29 stained in HeLa cells. Cells were fixed with 100% methanol (5min) 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-specificprotein-protein interactions. The cells were then incubated with theantibody 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) andab150080 (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.
Immunocytochemistry/ Immunofluorescence - AntiPCNA antibody [PC10] (ab29)

Overlay histogram showing HeLa cells stained with ab29 (red line).
The cells were fixed with 80% methanol (5 min) and thenpermeabilized with 0.1% PBS-Tween 20 for 20 min. The cells werethen incubated in 1x PBS / 10% normal goat serum / 0.3M glycine
to block non-specific protein-protein interactions followed by theantibody (ab29, 0.1μg/1x106 cells) for 30 min at 22ºC. The
secondary antibody used was Alexa Fluor® 488 goat anti-mouse
IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22ºC. Isotypecontrol 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.
Flow Cytometry (Intracellular) - Anti-PCNA antibody

Acquisition of >5,000 events were collected using a 20mW Argon[PC10] (ab29)

ion laser (488nm) and 525/30 bandpass filter.

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ab29 at 1/6000 staining mouse embryo (day 17) liver and gut tissue
sections by IHC-P. The tissue was formaldehyde fixed and a heat
mediated antigen retrieval step in Tris buffer was performed. Thetissue was blocked before incubation with the antibody for 2 hours.
A biotinylated goat polyclonal antibody was used as the secondary.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-PCNA antibody [PC10](ab29)
This image is courtesy of an abreview submitted byCarl Hobbs (King's College London, United Kingdom)

Immunohistochemistical staining (Formaldehyde/PFA-fixedparaffin-embedded sections) for PCNA antibody [PC10] Proliferation Marker (ab29) on Rat Tissue sections (adult spinalcord DRG). Antigen retrieval step: Heat mediated. Blocking step:
1% BSA for 10 mins at RT. Primary Antibody used at 1/6000 for 2
minutes at RT. Secondary Antibody: Biotin labelled goat anti mouseIgs (1/200).

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-PCNA antibody [PC10](ab29)
This image is courtesy of Carl Hobbs, King's CollegeLondon, United Kingdom

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Mouse monoclonal [PC10] to PCNA - Proliferation Marker (ab29)
used in immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections; 1/6000 for 2h at RT) on E6 developing chickbrain). Antigen retrieval step: Heat mediated. Blocking step: 1%
BSA for 10 mins at RT. Incubation time: Secondary Antibody: Biotin
conjugated goat anti mouse Igs (1/200). NB: This image showsdeveloping brain/overlying skin.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-PCNA antibody [PC10](ab29)
This image is courtesy of Carl Hobbs, King's CollegeLondon, United Kingdom

Mouse monoclonal [PC10] to PCNA - Proliferation Marker (ab29)
used in immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections;1/6000 for 2h at RT) on intestine of adultzebrafish). Antigen retrieval step: Heat mediated. Blocking step: 1%
BSA for 10 mins at RT. Incubation time: Secondary Antibody: Biotin
conjugated goat anti mouse Igs (1/200). NB: The crypt nuclei on thisimage of zebrafish intestine, are positive for the PCNA/PC10 clone
conforming to accepted localisation data for PCNA in otherspecies.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-PCNA antibody [PC10](ab29)
This image is courtesy of Carl Hobbs, King's CollegeLondon, United Kingdom

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IHC image of PCNA staining in rat large intestine formalin fixedparaffin embedded tissue section, performed on a Leica Bondsystem using the standard protocol F. The section was pre-treatedusing heat mediated antigen retrieval with sodium citrate buffer(pH6, epitope retrieval solution 1) for 20 mins. The section was thenincubated with ab29, 0.025µg/ml, for 15 mins at room temperatureand detected using an HRP conjugated compact polymer system.
DAB was used as the chromogen. The section was thencounterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated)customers should optimize variable parameters such as antigen
Immunohistochemistry (Formalin/PFA-fixed paraffin-retrieval conditions, primary antibody concentration and antibody

embedded sections) - Anti-PCNA antibody [PC10]

incubation times.

(ab29)

IHC image of PCNA staining in rat spleen formalin fixed paraffin
embedded tissue section, performed on a Leica Bond systemusing the standard protocol F. The section was pre-treated using
heat mediated antigen retrieval with sodium citrate buffer (pH6,epitope retrieval solution 1) for 20 mins. The section was thenincubated with ab29, 1/30,000 dilution 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.
For other IHC staining systems (automated and non-automated)customers should optimize variable parameters such as antigen
Immunohistochemistry (Formalin/PFA-fixed paraffin-retrieval conditions, primary antibody concentration and antibody

embedded sections) - Anti-PCNA antibody [PC10]

incubation times.

(ab29)

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Aldehyde dehydrogenase (ALDH1) expression correlateswith clinical outcome of breast cancer patients(A) Immunohistochemical staining shows tumors with poor clinicalresponse (progressive or stable disease, PD/SD) to neo-adjuvant
chemotherapy express high ALDH1 (>20% positive cancer cells) in
pre-chemotherapy samples, and tumors with partal response (PR)express low ALDH1 (≤20% positive cancer cells). High proliferatingcell nuclear antigen (PCNA)(>25% positive cancer cells) and poor
apoptosis are observed in tumors with PD/SD after neo-adjuvant
chemotherapy. Representative images of ALDH1 (×200), PCNAImmunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-PCNA antibody [PC10](×200) and Terminal deoxynucleotidyl transferase (TdT)-mediateddUTP labeling (TUNEL,×400).

(ab29)
Gong, C. et al PLoS One. 2010 Dec 20;5(12):e15630.
doi: 10.1371/journal.pone.0015630 Reproduced under
the Creative Commons license
http://creativecommons.org/licenses/by/4.0/

PCNA is detected using ab29 at 1/100 dilution.
(From Figure 1A of Gong et al)

Annexin V and PCNA staining of decellularized lungscaffolds recellularized with (A) MSCs in static versus
bioreactor conditions for 14 (panels A, B for annexin V, and a,b for PCNA) and 28 days (panels C, D for annexin V, and c, dfor PCNA) (single cells), (B) MSC cell clusters in static
conditions at 14 days (panel A for annexin V, and a for
PCNA), (C) C10 cells in static (panel A for annexin V, a forPCNA) versus bioreactor (panel B for annexin V, b for PCNA)
conditions for 11 days.
An inset in Fig 4Ab with higher magnification is shown todemonstrate that a majority of the cells stained positive for PCNA.
Immunohistochemistry (Formalin/PFA-fixed paraffin-

Cell nuclei are labeled in blue; marker of interest is labeled in green.

embedded sections) - Anti-PCNA antibody [PC10]

Magnifications are 400x. Overlap of cell nucleus and marker of(ab29)

interest can generate green or white color. For each condition,

Crabbe et al PLoS One. 2015 May 11;10(5):e0126846.
doi: 10.1371/journal.pone.0126846. eCollection 2015
Reproduced under the Creative Commons license
http://creativecommons.org/licenses/by/4.0/

images are representative of the entire lung.
MSCs = Bone marrow-derived mouse mesenchymal stromal (stem)
cells.
PCNA is detected using ab29 at 1/1000 dilution.
(From Figure 4 of Crabbe et al)

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