Rabbit Anti-PCNA antibody |
反应物种(预测) |
Rat,Chicken,Pig,Cow,Rabbit,Sheep,GuineaPig |
产品应用(已验证) |
WB,IHC,IF,FCM |
产品应用(可尝试) |
ELISA |
推荐稀释比例 |
WB=1:500-2000,Elisa=1:5000-10000,IHC-P=1:100-500,IHC-F=1:100-500,Flow Cyt=1ug/Test,IF=1:100-500, |
研究领域 |
肿瘤,细胞生物,免疫学,染色质和核信号,细胞周期蛋白,转录调节因子,细胞类型标志物(Proliferation Marker), |
标签 |
Array |
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Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6) 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) Polyclonal Antibody, Unconjugated (bs-2006R) at 1:200 overnight at 4°C, followed by a conjugated secondary (bs-0295G-Cy3) at [1:500] for 90 minutes and DAPI staining of the nuclei.
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Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6) 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) Polyclonal Antibody, Unconjugated (bs-0754R) at 1:200 overnight at 4°C, followed by a conjugated secondary (bs-0295G-Cy3) at [1:500] for 90 minutes and DAPI staining of the nuclei.
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Tissue/cell: human glioma tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-PCNA Polyclonal Antibody, Unconjugated(bs-2006R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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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) Polyclonal Antibody, Unconjugated (bs-2006R) at 1:200 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
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Blank control (blue line): U251 (blue).
Primary Antibody (green line): Rabbit Anti-PCNA antibody (bs-2006R)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min)and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 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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Sample:Spleen (Mouse) Lysate at 40 ug
Primary: Anti-PCNA (bs-2006R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 29 kD
Observed band size: 32 kD
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Blank control:Jurkat.
Primary Antibody (green line): Rabbit Anti-PCNA antibody (bs-2006R)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/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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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) Polyclonal Antibody, Unconjugated (bs-2006R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Sample:
Lane 1: Hela (Human) Cell Lysate at 30 ug
Lane 2: MCF-7 (Human) Cell Lysate at 30 ug
Lane 3: MOLT-4 (Human) Cell Lysate at 30 ug
Lane 4: HepG2 (Human) Cell Lysate at 30 ug
Primary: Anti-PCNA (bs-2006R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 32 kD
Observed band size: 32 kD
RRID:AB_10855815
产品名称:Rabbit Anti-PCNA antibody
别名: Proliferation Marker; Cyclin; DNA polymerase delta auxiliary protein; HGCN8729; MGC8367; Mutagen-sensitive 209 protein; Pcna/cyclin; PCNAR; Polymerase delta accessory protein; Proliferating Cell Nuclear Antigen; PCNA_HUMAN.
中文名称:增殖细胞核抗原抗体
英文名称:Rabbit Anti-PCNA antibody
抗体来源: Rabbit
克隆类型:多克隆
细胞定位:细胞核
性 状:Liquid
亚 型:IgG
纯化方法:affinity purified by Protein A
保存条件:Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
免 疫 原:KLH conjugated synthetic peptide derived from human PCNA
抗原表位:151-261/261
SWISS:P12004
Gene ID :5111
Human Gene ID:5111
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.
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
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 misi
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.
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. Follo
Similarity:Belongs to the PCNA family.
Important Note:This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.