Cell proliferation was quantified using Ki67 immunostaining. Briefly, HRMECs with different transfection factors were fixed in 4% paraformaldehyde (PFA; Beyotime, Jiangsu, China), followed by permeabilization with 0.2% Triton-X in PBS (Beyotime). After blocking with 10% goat serum for 30 minutes, the cells were incubated with a mouse anti-Ki67 antibody (Boster Biological Technology Co., Ltd., Wuhan, China) overnight at 4°C, followed by incubation with goat anti-mouse IgG-Alexa 594 (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA) for one hour at room temperature. The cells were observed in six randomly selected fields under a fluorescence microscope (BX63; Olympus, Tokyo, Japan) after nuclear staining with 4’,6-diamidino-2-phenylindole (Sigma-Aldrich, St. Louis, MO, USA), and the ratio of Ki67-positive cells to the number of total cells was calculated.
Mouse anti ki67 antibody
Manufactured by Boster Bio
Sourced in China
The Mouse anti-Ki67 antibody is a laboratory reagent used for the detection and quantification of the Ki67 protein, a cellular marker associated with proliferation. It is designed for use in various immunoassay techniques, such as immunohistochemistry and flow cytometry, to identify and analyze Ki67-positive cells.
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2 protocols using mouse anti ki67 antibody
1
Quantification of Cell Proliferation via Ki67 Immunostaining
2
Protein Expression Analysis of Colorectal Cancer Cell Lines
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After transfection, SW480 and HCT116 cells were collected, followed by protein extraction using RIPA lysis buffer. After 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis, isolated proteins were electro-transferred to polyvinylidene fluoride membranes and then blocked for 60 min with 5% skim milk. Subsequently, the membranes were incubated with primary antibodies from Boster (Wuhan, China) at 4 °C overnight, followed by incubation with horseradish peroxidase-conjugated secondary goat antibody (catalog number: BA1056) from Boster for 1 h at room temperature. The primary antibodies used in this experiment were list as follows: mouse anti-Ki-67 antibody (catalog number: M00254-1, Boster), mouse anti-proliferating cell nuclear antigen antibody (PCNA, catalog number: BM0104, Boster), rabbit anti-β-actin antibody (catalog number: BM3873, Boster), rabbit anti-E-cadherin antibody (catalog number: M00063-3, Boster), rabbit anti-Vimentin antibody (catalog number: BM4029, Boster), mouse anti-N-cadherin antibody (catalog number: BM1573, Boster).
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