Cells were fixed, permeabilized, blocked against nonspecific adhesion, immunostained for target proteins, and then imaged on an inverted Eclipse Ti epifluorescence microscope (Nikon). Primary antibodies were administered at manufacturer recommended concentration: anti-human CD45 (clone HI30, Becton Dickinson), anti-mouse CD45 (clone 30-F11, Becton Dickinson), anti-human mitochondria (clone 113–1, MilliporeSigma), anti-Ki67 (clone 8D5, Cell Signaling Technology), and anti-phospho-histone H2A.X (Ser139, clone 20E3, Cell Signaling Technology). The secondary antibodies used were Alexa Fluor 488 goat anti-mouse IgG (H+L) (Invitrogen) and Alexa Fluor 568 goat anti-rabbit IgG (H+L) (Invitrogen).
Anti ki67 clone 8d5
Manufactured by Cell Signaling Technology
Sourced in Singapore
Anti-Ki67 (clone 8D5) is a mouse monoclonal antibody that recognizes the Ki-67 protein, a cellular marker associated with cell proliferation. The antibody is designed for use in immunohistochemical applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti human cd45 clone hi30, by BD (2 mentions)
Anti mouse cd45 clone 30 f11, by BD (2 mentions)
Alexa fluor 568 goat anti rabbit igg h l, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488 goat anti mouse igg h l, by Thermo Fisher Scientific (2 mentions)
Anti human mitochondria clone 113 1, by Merck Group (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Anti αsma clone 1a4, by Merck Group (1 mentions)
Clone 8d5, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Ebioscience intracellular fixation permeabilization buffer set, by Thermo Fisher Scientific (1 mentions)
Muse cell analyzer, by Merck Group (1 mentions)
3 protocols using anti ki67 clone 8d5
1
Immunofluorescence Staining of Cells
2
Immunofluorescence Staining of Cells
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Cells were fixed, permeabilized, blocked against nonspecific adhesion, immunostained for target proteins, and then imaged on an inverted Eclipse Ti epifluorescence microscope (Nikon). Primary antibodies were administered at manufacturer recommended concentration: anti-human CD45 (clone HI30, Becton Dickinson), anti-mouse CD45 (clone 30-F11, Becton Dickinson), anti-human mitochondria (clone 113–1, MilliporeSigma), anti-Ki67 (clone 8D5, Cell Signaling Technology), and anti-phospho-histone H2A.X (Ser139, clone 20E3, Cell Signaling Technology). The secondary antibodies used were Alexa Fluor 488 goat anti-mouse IgG (H+L) (Invitrogen) and Alexa Fluor 568 goat anti-rabbit IgG (H+L) (Invitrogen).
3
miR-221 Regulates Cell Proliferation
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miR-221 mimics or control (MC) were transfected at 25 nM using lipofectamine RNAiMax (Thermo Fisher Scientific, Singapore). Ki67 proliferation assay was performed 24 h after transfection. Briefly, cells were trypsinized, fixed, and permeabilized (eBioscience Intracellular Fixation & Permeabilization Buffer Set, Thermo Fisher Scientific, Singapore) for anti-ki67 Ab staining (clone 8D5, Cell Signaling Technology, Research Biolabs, Singapore) and anti-mouse IgG-PE (Invitrogen, Thermo Fisher Scientific, Singapore), followed by flow cytometry on MUSE cell analyzer (Merck, Singapore). Cell number was assessed on MUSE.
In vivo cell proliferation was assessed in MId7 heart cross sections by immunofluorescence staining with anti-ki67 (clone 8D5, Cell Signaling Technology, Research Biolabs, Singapore), anti-α-SMA (clone 1A4, Sigma-Aldrich, Merck, Singapore), and DAPI (4′,6-diamidino-2-phenylindole) (Invitrogen, Thermo Fisher Scientific, Singapore).
In vivo cell proliferation was assessed in MId7 heart cross sections by immunofluorescence staining with anti-ki67 (clone 8D5, Cell Signaling Technology, Research Biolabs, Singapore), anti-α-SMA (clone 1A4, Sigma-Aldrich, Merck, Singapore), and DAPI (4′,6-diamidino-2-phenylindole) (Invitrogen, Thermo Fisher Scientific, Singapore).
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