Retinal sections on slides were washed in PBS to remove O.C.T. compound and incubated in 1:1000 rabbit anti-GFP antibody (catalog no. A-11122; ThermoFisher) in PBS + 0.3% Triton X-100 overnight at RT. Antibody labeling was detected the following day by washing in PBS (3 × 10 minutes), incubating for 2 hours in 1:500 Alexa Fluor 488-conjugated donkey anti-rabbit secondary antibody (catalog no. 711-545-152; Jackson ImmunoResearch Laboratories, Bar Harbor, ME), washing again in PBS (3 × 10 minutes), and mounting for observation in Fluoroshield with 4′,6-diamidino-2-phenylindole (DAPI) histology mounting medium (catalog no. F6057-20ML; Sigma-Aldrich). For labeling of retinal whole mounts, retinas were equilibrated in PBS, then immersed in 1:500 rabbit anti-GFP antibody in PBS + 0.3% Triton X-100 for 1 to 3 days, washed in PBS (3 × 20 minutes), and incubated in 1:500 Alexa Fluor 488-conjugated donkey anti-mouse secondary overnight at RT. Whole mounts were then extensively washed in PBS (3 × 30 minutes) and mounted on slides with Fluoroshield with DAPI, flanked by coverslip spacers to avoid damage from excessive flattening when applying the coverslip.
Fluoroshield with 4 6 diamidino 2 phenylindole dapi histology mounting medium
Manufactured by Merck Group
Sourced in Montenegro
Fluoroshield with 4',6-diamidino-2-phenylindole (DAPI) is a histology mounting medium. It is designed for use in fluorescence microscopy applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using fluoroshield with 4 6 diamidino 2 phenylindole dapi histology mounting medium
1
GFP Immunohistochemistry of Retinal Tissue
2
Immunofluorescence Analysis of Colonic Tissue
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Colonic tissues were frozen in optimal cutting temperature compound (OCT) cryomold in liquid nitrogen. 5μm sections were cut and fixed with ice-cold acetone. The sections were then washed with PBS and blocked with 1% bovine serum albumin (BSA) and Avidin Biotin Blocking Kit (Abcam, Cambridge, England) according to the instructions. Then the tissue sections were incubated with primary antibodies including zonula occludens-1 (ZO-1, 1:200), E-cadherin (1:200) and leucine rich repeat containing G protein-coupled receptor 5 (Lgr5, 1:100). After overnight incubation at 4°C, secondary antibodies were incubated 1 h at room temperature followed by counterstained nuclei using Fluoroshield™ with 4′,6-diamidino-2-phenylindole (DAPI) histology mounting medium (Sigma-Aldrich) and examined by immunofluorescence microscopy (Olympus).
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