HeLa cells were cultured in RPMI medium minus arginine and lysine supplemented with 10% dialysed foetal calf serum, and with 76 µg/ml arginine and 133 µg/ml lysine (either light R0K0 or heavy R10K8). Trypsin-free cell dissociation buffer from invitrogen was used for passaging cells. Cells were passaged nine times to ensure full incorporation of heavy amino acids which was checked by mass spectrometry to be >98% heavy label incorporation.
Trypsin free cell dissociation buffer
Manufactured by Thermo Fisher Scientific
Trypsin-free cell dissociation buffer is a cell culture reagent designed to gently detach adherent cells from the culture surface without the use of trypsin. It is a buffered, sterile-filtered solution that helps maintain cell viability and function during the cell harvesting process.
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Lab products found in correlation
2 protocols using trypsin free cell dissociation buffer
1
SILAC Labeling of HeLa Cells
2
Characterization of Wnt Receptor Expression
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Cells were collected using trypsin-free cell dissociation buffer (Invitrogen) and resuspended in FACS buffer (PBS with 1% BSA and 0.02% sodium azide). After blocking with mouse serum, cells were incubated with anti-FZD6 or anti-LRP6 antibody (R&D system) for 1 h at 4°C, followed by incubation with conjugated secondary antibodies. After washing in FACS buffer, cells were subjected to multichannel analysis using a FACScan flow cytometer (BD) and analyzed using Cell-Quest 3.3 software.
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