| 產(chǎn)品編號(hào) | bsm-33035M |
| 英文名稱 | Mouse Anti-PCNA(Nuclear Loading Control) antibody |
| 中文名稱 | 增殖細(xì)胞核抗原（內(nèi)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 |
| 產(chǎn)品類型 | 內(nèi)參抗體 |
| 研究領(lǐng)域 | 腫瘤 細(xì)胞生物 細(xì)胞周期蛋白 轉(zhuǎn)錄調(diào)節(jié)因子 細(xì)胞類型標(biāo)志物 |
| 抗體來(lái)源 | Mouse |
| 克隆類型 | Monoclonal |
| 克 隆 號(hào) | 10E9 |
| 交叉反應(yīng) | Rat,Mouse,Human |
| 產(chǎn)品應(yīng)用 | WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, Flow-Cyt=1:50 not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. |
| 理論分子量 | 29kDa |
| 細(xì)胞定位 | 細(xì)胞核 |
| 性 狀 | Liquid |
| 濃 度 | 1mg/ml |
| 免 疫 原 | KLH conjugated synthetic peptide derived from human PCNA |
| 亞 型 | IgG |
| 純化方法 | affinity purified by Protein G |
| 緩 沖 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. |
| 注意事項(xiàng) | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| PubMed | PubMed |
| 產(chǎn)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. 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. 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). 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. 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. Similarity: Belongs to the PCNA family. SWISS: P12004 Gene ID: 5111 Database links: Entrez Gene: 5111 Human Entrez Gene: 18538 Mouse Omim: 176740 Human SwissProt: P12004 Human SwissProt: P17918 Mouse Unigene: 147433 Human Unigene: 728886 Human Unigene: 7141 Mouse Unigene: 223 Rat PCNA是一種僅在增殖細(xì)胞中合成或表達(dá)的核內(nèi)多肽，其表達(dá)和合成與細(xì)胞周期有關(guān)。主要表達(dá)于增殖細(xì)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(xì)胞增殖和其惡性程度的一種指標(biāo). |
| 產(chǎn)品圖片 |  Sample: NIH/3T3 Cell (Mouse) Lysate at 40 ug Primary: Anti-PCNA (bsm-33035M) 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-33035M) 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: A431 Cell (Human) Lysate at 40 ug Primary: Anti-PCNA (bsm-33035M) 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: Lane 1: MCF-7 (Human) Cell Lysate at 30 ug Lane 2: HepG2 (Human) Cell Lysate at 30 ug Lane 3: Molt-4 (Human) Cell Lysate at 30 ug Lane 4: NIH/3T3 (Mouse) Cell Lysate at 30 ug Primary: Anti- PCNA (bsm-33035M) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 36 kD Observed band size: 35 kD  Paraformaldehyde-fixed, paraffin embedded (mouse testis tissue); 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33035M) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0024) for 20 minutes and DAB staining.  Paraformaldehyde-fixed, paraffin embedded (Human colon carcinoma); Antigen retrieval by microwave in sodium citrate buffer (pH6.0) ; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30min; Antibody incubation with (PCNA) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP)and DAB staining.  Paraformaldehyde-fixed, paraffin embedded (Human stomach 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) Monoclonal Antibody, Unconjugated (bsm-33035M) 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) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0024) for 20 minutes and 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) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0024) for 20 minutes and DAB staining.  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) Monoclonal Antibody, Unconjugated (ascites of bsm-33035M) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0024) for 20 minutes and 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33035M) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0024) for 20 minutes and DAB staining.  Paraformaldehyde-fixed, paraffin embedded (rat colon tissue); 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33035M) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0024) for 20 minutes and DAB staining.  Paraformaldehyde-fixed, paraffin embedded (human rectal 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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (human tonsil); 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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (mouse spleen); 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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 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) Monoclonal Antibody, Unconjugated (bsm-33035M) 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 kidney); 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; Incubation with (PCNA(Nuclear Loading Control) ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (human colon); 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; Incubation with (PCNA(Nuclear Loading Control) ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (mouse lymphoid); 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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (human cervical 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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (Descending colon cancer); 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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (Human stomach 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) Monoclonal Antibody, Unconjugated (bsm-33035M) 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 cancer); 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 (bsm-33035M) 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 esophageal cancer); 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(Nuclear Loading Control) ) Monoclonal Antibody, Unconjugated (bsm-33035M ) at 1:200 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) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 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) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (human tonsil); 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) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (human cervical 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) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (rat lymphoid); 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) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (mouse colon); 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 ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 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 ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (rat colon); 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 ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 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 ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 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 ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (Mouse small intestine); 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 ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (rat kidney); 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 ) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (human cervical 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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Paraformaldehyde-fixed, paraffin embedded (rat spleen); 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(Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (bsm-33035M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.  Blank control:Jurkat. Primary Antibody (green line): Mouse Anti-PCNA antibody (bsm-33035M) Dilution: 1:50; Secondary Antibody : Goat anti-mouse 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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