1.105 cells were cultured at 37 °C in 5% CO2 for 48 h in a polystyrene vessel chamber on tissue culture treated glass slide (BD Falcon). Cells were fixed with 4% paraformaldehyde in PBS for 30 min and washed with PBS Ca2+/Mg2+ (Sigma-Aldrich). Cells were then permeabilized with 0.1% Triton-X-100 (Sigma-Aldrich) for 5 minutes, then incubated for 1 h with a saturation solution (10% of goat serum (Sigma-Aldrich), 3% of human serum (Sigma-Aldrich), 0.1% Triton-X-100 (Sigma-Aldrich), 1% BSA (Euromedex), 0.05% Tween 20 (Euromedex) in PBS. Cells were then incubated at 4 °C overnight with the anti-MRP1 monoclonal antibody MRPm6 (Alexis Biochemical), washed and then incubated for 1 h with goat secondary antibody anti-mouse IgG coupled to Alexa Fluor 488 (Life Technologies). Nuclei were counterstained with 4 µM of Hoechst 33258 for 10 min. Samples were examined under a Compact Confocal Power Pack (ZEISS LSM 800 with Airyscan) with a 40 (numerical aperture, 1.2) oil immersion lens. The magnification used may be different depending on the samples.
Tissue culture treated glass slide
Manufactured by BD
The tissue culture treated glass slide is a laboratory equipment item designed for cell and tissue culture applications. It serves as a substrate for the growth and maintenance of various cell types in vitro. The surface of the slide has undergone a specialized treatment to enhance cell adhesion and proliferation, providing an optimal environment for cell culture experiments and analyses.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Euromedex (1 mentions)
Tween 20, by Euromedex (1 mentions)
Pbsca2 mg2, by Merck Group (1 mentions)
Human serum, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
Anti ha, by Cell Signaling Technology (1 mentions)
Ix71 microscope, by Olympus (1 mentions)
Anti flag, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Vectashield antifade mounting medium, by Vector Laboratories (1 mentions)
Cy3 conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Biotium (1 mentions)
Anti lc3 antibody, by Cell Signaling Technology (1 mentions)
Fluoro gel, by Electron Microscopy Sciences (1 mentions)
Tcs sp5, by Leica (1 mentions)
3 protocols using tissue culture treated glass slide
1
Immunofluorescence Staining of MRP1
2
Immunofluorescence Analysis of Transfected Cells
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Transfected cells (10,000–30,000 cells/well) were seeded in a chamber with tissue culture-treated glass slide (Falcon, 354118). For immunofluorescence analysis, cells were fixed with 4% paraformaldehyde for 20 mins and then permeabilized with 1X PBS containing 0.1% Triton X-100. Cells were incubated with anti-FLAG (1:400 dilution; Cell Signaling, 14793), and anti-HA (1:100 dilution; Cell Signaling, 2367) overnight at 4°C. Alexa Fluor Plus 647- conjugated secondary antibodies (1:400 dilution; Invitrogen, A32733) and Cy3-conjugated secondary antibodies (1:1000 dilution; Jackson ImmunoResearch, 115-165-003) were added followed by incubation at 37°C for 30 mins. Cells were rinsed several times with 1X PBS, stained with DAPI (1 μg/mL; Invitrogen, 62247), and mounted with Vectashield Antifade mounting medium (Vector Laboratories, Inc., H1000). Image acquisition was performed using Olympus Microscope IX71 or BZ-X810 All-in-One Fluorescent Microscopes.
3
Immunofluorescence Analysis of Autophagy
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The PROM1/CD133+ and PROM1/CD133− cells were plated on Tissue Culture Treated Glass Slide (BD Falcon, 354114) and subjected to PDT treatment (1.3 J/cm2). After 24 h, cells were washed with PBS and fixed with 3.7% paraformaldehyde for 8 min. Fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 15 min, blocked with 3% BSA for 60 min, and incubated with anti-LC3 antibodies (1:200) (Cell Signaling Technology, 3868S) overnight. After washing with PBS, cells were incubated for 1 h with DyLight 488 conjugated secondary antibody (1:500) (BioLink Biotechnology, B0210). Nuclei were counterstained with DAPI (Biotium, 40043) for 5 min. Slides were mounted with a coverslip using Fluoro-Gel (Electron Microscopy Sciences, 17985-10). Cells were imaged using a confocal laser scanning microscope (TCS SP5, Leica Microsystems, Wetzlar, Germany), and the images were analyzed using a Leica Application Suite 2.02. DyLight 488 was excited using an argon laser (488 nm), and DAPI was excited using a diode laser (405 nm). The emission lights for DAPI and DyLight 488 were collected between 410 and 490 nm and between 495 and 540 nm, respectively.
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