?英文名稱 PCNA(Nuclear Loading Control)?
中文名稱 增殖細胞核抗原（核內參）單克隆抗體?
別? ? 名 Cyclin; DNA polymerase delta auxiliary protein; HGCN8729; MGC8367; Mutagen-sensitive 209 protein; Pcna/cyclin; PCNAR; Polymerase delta accessory protein; Proliferating Cell Nuclear Antigen.? ?
研究領域腫瘤? 細胞生物? 免疫學? 細胞周期蛋白? 轉錄調節因子??
抗體來源Mouse
克隆類型Monoclonal
克 隆 號1C11
交叉反應Human, Mouse, Rat,? (predicted: Chicken, Cow, Rabbit, )
產品應用WB=1:400-1000 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1ug/Test IF=1:100-500 （石蠟切片需做抗原修復）
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
分 子 量29kDa
細胞定位細胞核?
性? ? 狀Liquid
濃? ? 度1mg/ml
免 疫 原KLH conjugated synthetic peptide derived from human PCNA:151-261/261?
亞? ? 型IgG
純化方法affinity purified by Protein G
儲 存 液0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存條件Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
PubMedPubMed
產品介紹Proliferating cell nuclear antigen (PCNA) is a 28kDa nuclear protein associated with the cell cycle, a nuclear protein vital for cellular DNA synthesis. Proliferating cell nuclear antigen was originally identified by immunofluorescence as a nuclear protein whose appearance correlated with the proliferate state of the cell. PCNA is required for replication of DNA in vitro and has been identified as the auxiliary protein (cofactor) for DNA polymerase delta. The anti-PCNA antibodies react with the nuclei of proliferating cells. PCNA is essential for cellular DNA synthesis and is also required for the in vitro replication of simian virus 40 (SV40) DNA where it acts to coordinate leading and lagging strand synthesis at the replication fork. The PCNA protein may fulfil several separate roles in the cell nucleus associated with changes in its antigenic structure.
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Function:
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. 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. 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.
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Subunit:
Homotrimer. Forms a complex with activator 1 heteropentamer in the presence of ATP. Interacts with EXO1, POLH, POLK, DNMT1, ERCC5, FEN1, CDC6 and POLDIP2. Interacts with APEX2; this interaction is triggered by reactive oxygen species and increased by misincorporation of uracil in nuclear DNA. Forms a ternary complex with DNTTIP2 and core histone. Interacts with KCTD10 and PPP1R15A (By similarity). Interacts with POLD1, POLD3 and POLD4. Interacts with BAZ1B; the interaction is direct. Interacts with HLTF and SHPRH. Interacts with NUDT15. Interaction is disrupted in response to UV irradiation and acetylation. Interacts with CDKN1A/p21(CIP1) and CDT1; interacts via their PIP-box which also recruits the DCX(DTL) complex. Interacts with DDX11. Interacts with EGFR; positively regulates PCNA. Interacts with PARPBP. Interacts (when ubiquitinated) with SPRTN; leading to enhance RAD18-mediated PCNA ubiquitination. Interacts (when polyubiquitinated) with ZRANB3. Interacts with SMARCAD1. Interacts with CDKN1C. Interacts with KIAA0101/PAF15 (via PIP-box).
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Subcellular Location:
Nucleus. Note=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.
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Post-translational modifications:
Following DNA damage, can be either monoubiquitinated to stimulate direct bypass of DNA lesions by specialized DNA polymerases or polyubiquitinated to promote recombination-dependent DNA synthesis across DNA lesions by template switching mechanisms. Following induction of replication stress, monoubiquitinated by the UBE2B-RAD18 complex on Lys-164, leading to recruit translesion (TLS) polymerases, which are able to synthesize across DNA lesions in a potentially error-prone manner. An error-free pathway also exists and requires 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. This error-free pathway, also known as template switching, employs recombination mechanisms to synthesize across the lesion, using as a template the undamaged, newly synthesized strand of the sister chromatid. 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. Sumoylated during S phase.
Acetylated in response to UV irradiation. Acetylation disrupts interaction with NUDT15 and promotes degradation.
Phosphorylated. Phosphorylation at Tyr-211 by EGFR stabilizes chromatin-associated PCNA.
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Similarity:
Belongs to the PCNA family.
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SWISS:
P12004
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Gene ID:
5111
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Database links:
Entrez Gene: 515499 Cow
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Entrez Gene: 5111 Human
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Entrez Gene: 18538 Mouse
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Entrez Gene: 25737 Rat
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Omim: 176740 Human
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SwissProt: Q3ZBW4 Cow
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SwissProt: P12004 Human
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SwissProt: P17918 Mouse
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SwissProt: P04961 Rat
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Unigene: 147433 Human
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Unigene: 728886 Human
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Unigene: 7141 Mouse
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Unigene: 223 Rat
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Important Note:
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
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PCNA是一種僅在增殖細胞中合成或表達的核內多肽，其表達和合成與細胞周期有關。主要表達于增殖細胞的S期、G1期和G2初期。
PCNA主要作為判斷各種惡性腫瘤(包括胃腸道癌腫、乳腺癌、肝癌、膀胱癌等)細胞增殖和其惡性程度的一種指標.
產品圖片
Sample: A431 Cell (Human) Lysate at 40 ug
Primary: Anti-PCNA (bsm-2006M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 29 kD
Observed band size: 29 kD
Sample:
Hela Cell (Human) Lysate at 40 ug
Primary: Anti-PCNA (bsm-2006M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 29 kD
Observed band size: 29 kD
Sample: NIH/3T3 Cell (Mouse) Lysate at 40 ug
Primary: Anti-PCNA (bsm-2006M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 29 kD
Observed band size: 29 kD
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (pcna(1c11)) Monoclonal Antibody, Unconjugated (bsm-2006M) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (pcna(1c11)) Monoclonal Antibody, Unconjugated (bsm-2006M) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (pcna(1c11)) Monoclonal Antibody, Unconjugated (bsm-2006M) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (pcna(1c11)) Monoclonal Antibody, Unconjugated (bsm-2006M) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (pcna(1c11)) Monoclonal Antibody, Unconjugated (bsm-2006M) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human liver carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (pcna(1c11)) Monoclonal Antibody, Unconjugated (bsm-2006M) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Blank control:Molt4.
Primary Antibody (green line): Mouse Anti-PCNA(1C11) antibody (bsm-2006M)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Mouse IgG .
Secondary Antibody : Goat anti-mouse IgG-CY5
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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