Twenty-four hours after transfection, the ACHN cells cultured on glass coverslips were fixed in 4% paraformaldehyde for 15 min and permeabilized with 0.01% Triton X-100 for 20 min at room temperature. After that, we had them blocked with 5% goat serum for 1 h at 37°C and performed an incubation with primary antibodies overnight at 4°C. The secondary antibodies were Alexa 488-conjugated goat anti-mouse or anti-rabbit IgG (Abcam, UK). Implementation of 4′,6-diamidino-2-phenylindole (Sigma-Aldrich) was for counterstaining the nuclei. Then, we took out fluorescent images with fluorescence microscope (IX70, Olympus, Japan). The primary antibodies were purchased from Abcam as follows: anti-LC3B (no. ab225383, 1 : 1000), anti-TRAF6 (no. ab40675, 1 : 500), anti-TP53INP2 (no. ab273012, 1 : 100), and caspase-8 (no. ab25901, 1 : 100).
Alexa 488 conjugated goat anti mouse or anti rabbit igg
Manufactured by Abcam
Sourced in United Kingdom
Alexa Fluor 488-conjugated goat anti-mouse or anti-rabbit IgG is a secondary antibody that binds to mouse or rabbit primary antibodies. It is labeled with the Alexa Fluor 488 fluorescent dye, which can be detected using standard FITC filter settings.
Automatically generated - may contain errors
2 protocols using alexa 488 conjugated goat anti mouse or anti rabbit igg
1
Immunofluorescence Analysis of Autophagy and Apoptosis Markers
2
Immunofluorescence Analysis of Active β-catenin and Snail1
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At 24 h following transfection, the cells cultured on glass coverslips were fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.01% Triton X-100 for 20 min at room temperature, blocked with 5% goat serum for 1 h at 37°C, and incubated with primary antibodies against non-phospho (active) β-catenin (CST, 19,807, 1:100) and Snail1 (Affinity, AF6032, 1:100) overnight at 4°C. The secondary antibodies were Alexa 488-conjugated goat anti-mouse or anti-rabbit IgG (Abcam). 4′,6-diamidino-2-phenylindole (10 µg/mL, 32,670; Sigma-Aldrich) was used to counterstain the nuclei. Fluorescent images were obtained under a microscope (Carl Zeiss, Germany) and processed using Photoshop software (Adobe).
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