Anti-PCNA Antibody Picoband®

CircSETD3 inhibits the growth of HCC through thecircSETD3/miR-421/MAPK14 pathway. a and b Both qRT-PCR and IHC showed MAPK14 was significantly downregulated in HCC tissues compared with matched non-tumorous tissues. c and d MAPK14 negatively correlated with miR-421 whereas positively correlated with circSETD3 in HCC tissues. e Schematic of MAPK14 wild-type (WT) and mutant (Mut) luciferase reporter vectors. f The relative luciferase activities were analyzed in 293 T cells co-transfected with miR-421 mimics or miR-mimics-NC and WT or Mut luciferase reporter vectors. g MiR-421 inhibitor up-regulated MAPK14 and down-regulated cyclinD1 and PCNA in Hep3B cells. MiR-421 mimics down-regulated MAPK14 and up-regulated cyclinD1 and PCNA in Huh7 cells. h CircSETD3 letivirus up-regulated MAPK14 and down-regulated cyclinD1 and PCNA in Huh7 cells, this effect could be reversed by co-transfected with miR-421 mimics. i circSETD3 siRNA down-regulated MAPK14 and up-regulated cyclinD1 and PCNA in Hep3B cells, this effect can be reversed by co-transfected with miR-421 inhibitors. HCC, hepatocellular carcinoma; qRT-PCR, quantitative reverse transcription polymerase chain reaction; in, inhibitors; mi, mimics; IHC, immunohistochemistry. ***P < 0.001. Error bars indicate SD
Index in PubMed under a CC BY license. PMID: 30795787

CircSETD3 stably maintained in xenograft tumor models and inhibit tumor growth by targeting MAPK14. a and b Smaller tumor size and lower tumor weight were observed in circSETD3-overexpressing group. c Over-expressed circSETD3 could stably maintained in xenograft tumor models. d The expression of MAPK14 was increased, Ki-67, PCNA and Cyclin D1 were decreased in circSETD3-overexpressing group when compared with control group. **P < 0.01, ***P < 0.001. Error bars indicate SD
Index in PubMed under a CC BY license. PMID: 30795787

Effects of each treatment on the growth of human OS xenograft tumors. Nude mice bearing 143 human OS xenograft tumors were treated with CAP (20 mg/kg) and DDP (4 mg/kg) alone or in combination. The volumes of the xenograft tumors were measured at the indicated time points ( a ). The weight of each nude was measured at the indicated time points ( b ). After the last treatment, the mice were sacrificed, and representative images of the subcutaneous tumor xenografts in the nude mice and the morphology of the tumors are presented ( c ). Tumors were collected, and the tumor weights were measured and compared ( d ). Xenograft tumors were sectioned and stained with PCNA and Ki67 via IHC ( e ). Statistical analyses of the expression of PCNA and Ki67 in different groups ( f ). Tumors were sectioned and stained with H&E, and representative histopathological images are presented ( g ). Effects of the different treatments on renal histology. Representative histological profiles of kidneys after the different treatments were detected by H&E staining ( h ). Effects of the different treatments on BUN and creatinine levels in mice ( i ). The quantitative data are shown as the mean ± SD of 5 independent experiments; * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. the control (CAP-, DDP-)
Index in PubMed under a CC BY license. PMID: 30326933

Western blot analysis of PCNA using anti-PCNA antibody (A00125).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human 293T whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: mouse NIH/3T3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PCNA antigen affinity purified polyclonal antibody (Catalog # A00125) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PCNA at approximately 36 kDa. The expected band size for PCNA is at 29 kDa.

IHC analysis of PCNA using anti-PCNA antibody (A00125).
PCNA was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-PCNA Antibody (A00125) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of PCNA using anti-PCNA antibody (A00125).
PCNA was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-PCNA Antibody (A00125) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of PCNA using anti-PCNA antibody (A00125).
PCNA was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-PCNA Antibody (A00125) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of PCNA using anti-PCNA antibody (A00125).
PCNA was detected in a paraffin-embedded section of human papillary thyroid carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-PCNA Antibody (A00125) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of PCNA using anti-PCNA antibody (A00125).
PCNA was detected in a paraffin-embedded section of human glioblastoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-PCNA Antibody (A00125) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of PCNA using anti-PCNA antibody (A00125).
PCNA was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-PCNA Antibody (A00125) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of PCNA using anti-PCNA antibody (A00125).
PCNA was detected in a paraffin-embedded section of rat colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-PCNA Antibody (A00125) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of PCNA using anti-PCNA antibody (A00125) and anti-Beta Tubulin antibody (M01857-3).
PCNA was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-PCNA Antibody (A00125) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of 293T cells using anti-PCNA antibody (A00125).
Overlay histogram showing 293T cells stained with A00125 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PCNA Antibody (A00125, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.