Cells were lysed in SDS lysis buffer (240 mmol/L Tris‐acetate, 1% SDS, 1% glycerol, 5 mmol/L EDTA pH 8.0) with DTT, protease inhibitors, and a cocktail of phosphatase inhibitors. Anti‐rabbit and anti‐mouse IgG secondary antibodies were from Invitrogen (Carlsbad, CA, USA) and LI‐COR BioSciences (Lincoln, NE, USA). Antibodies detecting AR, Snail, MMP‐2, and vimentin were purchased from Abcam (Abcam, Cambridge, UK). The antibody detecting E‐cadherin was purchased from BD BioSciences (BD BioSciences, San Jose, CA, USA). The antibodies detecting Slug and β‐catenin were purchased from Cell Signaling Technology (Cell Signaling Technology, Danvers, MA, USA). Antibody detecting KAT5 (Tip60) was from Abnova (Taipei, Taiwan) and GeneTex (Irvine, CA, USA). Antibody detecting CAV1 was from Millipore (Burlington, MA, USA). The antibody detecting β‐actin was purchased from Novus (Novus, Littleton, CO, USA).
Anti rabbit and anti mouse igg secondary antibodies
Manufactured by Thermo Fisher Scientific
Sourced in United States
Anti-rabbit and anti-mouse IgG secondary antibodies are laboratory reagents designed to detect and visualize primary antibodies raised in rabbit or mouse. These secondary antibodies are conjugated with various labels, such as fluorescent dyes or enzymes, to enable detection and quantification of target proteins in various immunoassays and immunohistochemistry applications.
Automatically generated - may contain errors
2 protocols using anti rabbit and anti mouse igg secondary antibodies
1
Immunoblotting Procedures for Epithelial-Mesenchymal Transition
2
miR-29a Regulates Cell Proliferation
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After 48 h of transfection with miR-29a mimics, cultures were stained with Hoechst dye and the number of cells in a fixed field was counted. For infection studies, supernatants of AAV2 viral particles either expressing miR-29a (titer: 2.82e + 13) a hardened siRNA-like mutant of miR-29a (titer: 3.71E + 13) or empty TR2 (titer: 2.15E + 13) vectors were incubated with the dermal fibroblasts for 72 h. Next, the cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 10 min at room temperature, rinsed in PBS, and processed for immunolabeling with rabbit anti-Ki67 (EnCor, Gainesville, FL) and mouse antivimentin (Santa Cruz) antibodies overnight at 4 °C. Bound primary antibodies were detected with anti-rabbit and anti-mouse IgG secondary antibodies (Invitrogen). Nuclei were stained with Hoechst dye (Invitrogen) and coverslips were mounted using ProLong Antifade mounting medium (Invitrogen). To determine the mitotic capacity of the cultures, Ki67-positive cells were counted in a fixed field (0.8 mm2), and divided by the total number of Hoechst dye positive cells. From each condition, the total number of cells in 6 to 170 randomly chosen fields were counted and graphed.
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