Computational protocol: Supporting a Role for the GTPase Rab7 in Prostate Cancer Progression

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Protocol publication

[…] I.F. microscopy was performed as previously described , . Briefly, cells were fixed with ice cold 4% PFA for 20minutes. Cells were then washed in PBS then incubated with anti-LAMP-1 (H4A3-s Iowa State University Hybridoma Bank) at a 1∶100 dilution in BSP for one hour at room temperature. Cells were then washed, and incubated with Dylight 594 anti mouse (Jackson IR) at a 1∶100 dilution in BSP for one hour at room temperature. Phalloidin was then used to stain the actin cytoskeleton (Phalloidin 488, Molecular Probes) and was incubated for 20 minutes at 1∶200 in BSP at room temperature. Slides were mounted with DAPI containing SlowFade gold reagent (Invitrogen S36938). Cells were imaged using an Olympus BX-50 microscope using MetaMorph software. Images were merged using ImageJ. […]

Pipeline specifications

Software tools MetaMorph, ImageJ
Application Microscopic phenotype analysis
Diseases Neoplasms, Prostatic Neoplasms