HeLa cells (60,000) were grown onto a coverslip (Coverglass for cell Growth™, Fisherbrand, USA) in a 24-well plate. The cells were treated with 6e , 6j , and 6n compounds at their IC50 dose for 12 h, 14 h, 16 h, and 18 h. Thereafter, cells were fixed in 4% paraformaldehyde (Himedia), washed with ice-cold 1X PBS, permeabilized with 0.25% Triton X-100 (Sigma), and washed with 0.1% PBST. The cell was counter stained with 49,6-diamidino-2-phenylindole, dihydrochloride (DAPI, Invitrogen) and mounted using one drop of ProLong® Gold antifade reagent (Invitrogen) on a slide. Images were acquired by fluorescence microscope (Leica SP8 TCS) and analyzed using ImageJ (NIH).
Coverglass for cell growth
Manufactured by Thermo Fisher Scientific
Sourced in United States
Coverglass for cell Growth™ is a laboratory equipment product specifically designed for cell culture applications. It provides a transparent surface for cells to adhere to and grow upon, facilitating microscopic observation and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Paraformaldehyde (pfa), by HiMedia (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by MP Biomedicals (1 mentions)
Pakt473, by Cell Signaling Technology (1 mentions)
2 protocols using coverglass for cell growth
1
Imaging Cytotoxic Compound Effects on HeLa Cells
2
Immunofluorescence Staining of pAKT in HUVECs
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60,000 HUVECs were seeded onto Coverglass for cell Growth (Fisherbrand) in 24-well plate. Cells were fixed in 4% paraformaldehyde (Himedia), washed with ice-cold 1X PBS, permeabilized with 0.25% Triton X-100 (Sigma), washed thrice with 0.1% PBST, and blocked in 5% BSA (MP Biochemicals) for 1 hour. Primary antibody (rabbit pAKT-473, dilution 1:100, Cell Signaling Technology, Cat#4060S) was added over the coverslips followed by incubation in a humidified chamber at 4 C for 16 hours. Furthermore, the coverslips were washed with 0.1% PBST and incubated with secondary antibody (goat anti-rabbit IgG (HþL) Alexa Fluor 594, dilution 1:500, Cat#A-11012, RRID:AB_141359) for 2 hours at room temperature followed by washing. Afterwards coverslips were counterstained with 4 0 ,6-diamidino-2-phenylindole, dihydrochloride (DAPI, Invitrogen, Cat#D1306) and mounted with one drop of ProLong Gold antifade reagent (Invitrogen, Cat#P36930) on the slide. Images were acquired by fluorescence microscope (Leica SP8) and analyzed using ImageJ (RRID:SCR_003070).
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