Anti-Ki67/MKI67 Antibody Picoband®

IHC analysis of Ki67 using anti-Ki67 antibody (PB9026).
Ki67 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Ki67 Antibody (PB9026) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Western blot analysis of Ki67 using anti-Ki67 antibody (PB9026).
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 50ug of sample under reducing conditions.
Lane 1: HELA Whole Cell Lysate,
Lane 2: MCF-7 Whole Cell Lysate,
Lane 3: COLO320 Whole Cell Lysate,
Lane 4: HEPG2 Whole Cell Lysate,
Lane 5: SKOV Whole Cell Lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Ki67 antigen affinity purified polyclonal antibody (Catalog # PB9026) 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 1:10000 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 Ki67 at approximately 358KD. The expected band size for Ki67 is at 358KD.

Therapeutic potentials of Pg-Fe-hMSC in both monocellular and multicellular humanized fibrotic models. a Schematic illustration and representative image of EpCAM immunostaining in primary human lung epithelial cells (hLECs). Scale bar, 50 μm. b Mitochondrial transfer rates from the indicated hMSC to the primary hLEC ( n = 3 biologically independent cells). c Relative intracellular ROS levels ( n = 3 biologically independent cells), d TGF- β expression levels ( n = 4 biologically independent cells), and e Viability of BLM-treated hLEC after the indicated treatment using different engineered hMSCs ( n = 3 biologically independent cells). f Schematic illustration showing the preparation of the 3D multicellular human fibrotic model. g Representative immunostaining images showing the expression of α -smooth muscle actin ( α -SMA), vimentin, collagen-I, and Ki-67 in the healthy and fibrotic multicellular human spheroid models. Blue fluorescent signals indicate the cell nuclei and green signals indicate the biomarkers. Scale bar, 20 μm. h Mitochondrial transfer rates of different engineered hMSC in fibrotic human lung spheroids ( n = 3 biologically independent experiments). i Relative intracellular ROS levels ( n = 3 biologically independent experiments) and j TGF- β expression levels of fibrotic human lung spheroids after the indicated treatment using different engineered hMSCs ( n = 4 biologically independent experiments). k Viability of fibrotic human lung spheroids after the indicated treatment using different engineered hMSCs ( n = 3 biologically independent experiments). Data are presented as means ± SD. Statistical significance was analyzed using ordinary one-way ANOVA. ECs epithelial cells.
Index in PubMed under a CC BY license. PMID: 37723135

IF analysis of Ki67 using anti-Ki67 antibody (PB9026).
Ki67 was detected in paraffin-embedded section of human colon organoid tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5μg/mL rabbit anti-Ki67 Antibody (PB9026) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

miR-134-3p overexpression inhibits HuOCSC activity both in vitro and in vivo . (A) Proliferation inhibition rate assay (n = 3). ** P < 0.01 vs. miR-mut. t test. (B) Cell cycle assay (n = 3). * P < 0.05 vs. miR-mut; ** P < 0.01 vs. miR-mut. t test. (C) Transwell assay of migration ability of HuOCSCs (n = 3). ** P < 0.01 vs. miR-mut. t test. (D) Angiogenesis assay to detect the ability of formation of tubular 3D structures (n = 3). ** P < 0.01 vs. miR-mut. t test. (E) In vitro graft tumor assay. * P < 0.05 vs. miR-mut. t test. (F) The weight and volume of graft tumors. * P < 0.05 vs. miR-mut. t test. (G) HE staining (2× magnification). (H) Immunofluorescence labeling of Ki67 (400× magnification).
Index in PubMed under a CC BY license. PMID: 38021163

ICC analysis of Ki67 using anti-Ki67 antibody (PB9026).
Ki67 was detected in immunocytochemical section of Hela cells. 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 2μg/ml rabbit anti-Ki67 Antibody (PB9026) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Silencing TFAP2A inhibits proliferation of TNBC cells. Expression of TFAP2A was knocked down by siRNAs in MDA-MB-231 and BT-549 cells, cell proliferation was assessed by A MTS assay, B Colony formation assay, and C EdU assay. D Western blot analysis showing the expression levels of Ki67, TFAP2A, and the phosphorylation levels of ERK and AKT in MDA-MB-231 cells and E BT-549 cells with or without TFAP2A knockdown
Index in PubMed under a CC BY license. PMID: 38890750

miR-8072 suppresses TNBC tumor progression and predicts favorable prognosis. ( A ) Serum levels of miR-8072 in breast cancer, benign breast disease, and non-cancer samples were analyzed using data from GEO (GSE73002) (** P < 0.01) . B The relationships between miR-8072 level and overall survival (OS) in breast cancer patients were analyzed using public database Kaplan–Meier plotter ( ). 1062 breast cancer patients were included, 525 cases were in miR-8072 low-expression cohort, and 537 cases were in miR-8072 high-expression cohort. The median survival for the high-expression cohort of miR-8072 is 115.4 months, while it is 148.53 months for the low-expression cohort of miR-8072 (upper panel). 97 TNBC patients were included for analysis. Among them, 28 cases were in miR-8072 low-expression cohort, and 69 cases were in miR-8072 high-expression cohort. The upper quartile survival for cohorts with high-expression of miR-8072 is 98.83 months, while it is 36.43 months for cohorts with low-expression of miR-8072 (lower panel). C MDA-MB-231 cells with stably overexpressed miR-8072 or control (Ctrl) cells were subcutaneously implanted in nude mice, tumor volume were measured every 7 days. D Tumor nodules were collected and weighed 35 days later after implantation. E Expression of Ki67 in the tumor nodules was detected by immunohistochemistry
Index in PubMed under a CC BY license. PMID: 38890750

IF analysis of Ki67 using anti-Ki67 antibody (PB9026).
Ki67 was detected in a paraffin-embedded section ofNude mouse tumor 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 1:200 rabbit anti-Ki67 Antibody (PB9026) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

ICC/IF analysis of Ki67 using anti-Ki67 antibody (PB9026).
Ki67 was detected in an immunocytochemical section of human Hela cells. The cells were fixed with 4% paraformaldehyde for 10 minutes and then treated with a membrane permeabilization agent (AR0205) for 5 minutes.The cells were blocked with 10% goat serum. And then incubated with rabbit anti-Ki67 Antibody (PB9026) at a dilution of 1:50 overnight at 4°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

ICC/IF analysis of Ki67 using anti-Ki67 antibody (PB9026).
Ki67 was detected in an immunocytochemical section of human SIHA cells. The cells were fixed with 4% paraformaldehyde for 10 minutes and then treated with a membrane permeabilization agent (AR0205) for 5 minutes.The cells were blocked with 10% goat serum. And then incubated with rabbit anti-Ki67 Antibody (PB9026) at a dilution of 1:50 overnight at 4°C. DyLight®488 conjugated goat Anti-rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of Ki67 using anti-Ki67 antibody (PB9026).
Ki67 was detected in paraffin-embedded section of nude mouse tumor tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:1000 rabbit anti-Ki67 Antibody (PB9026) overnight at 4°C. Ready-to-use SABC-POD kit (rabbit IgG) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.