Commercial human
uterine adenocarcinoma cancer cell line HEC-1-A (ATCC HTB-112) was
obtained from the American Type Culture Collection (ATCC, Manassas,
VA). HEC-1-A cells are cancer cells that use the telomerase enzyme
in their process of becoming immortal,36 (link) and telomerase is the target of our biosensor; thus, HEC-1-A represents
a positive control. To grow and subculture the HEC-1-A cell line,
we followed the manufacturer’s protocol37 and detailed it in theSupporting Information .
Telomerase extract preparation was accomplished through the
following steps:38 (link) (1) cells were collected
in the exponential phase of growth, and 1 × 106 cells/mL
were washed twice with ice-cold phosphate-buffered saline (PBS) 1×
(140 mM NaCl, 2.7 M KCl, 10 mM NaHPO4, and 1.8 mM KH2PO4). (2) Then, cells were resuspended in ice-cold
3-[3-(cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS)
lysis buffer [10 mM Tris–HCl, pH 7.5, 1 mM MgCl2, 1 mM ethylenediaminetetraacetic acid (EDTA), 0.1 mM phenylmethanesulfonyl
fluoride (PMSF), 0.5% CHAPS, 10% glycerol] with a final concentration
of 25 × 106 cells/mL. (3) The suspension was then
incubated for 30 min on ice and centrifuged for 20 min at 15,000 rpm,
4 °C. (4) Finally, the supernatant was carefully transferred
to a fresh tube and frozen at −80 °C until use.
uterine adenocarcinoma cancer cell line HEC-1-A (ATCC HTB-112) was
obtained from the American Type Culture Collection (ATCC, Manassas,
VA). HEC-1-A cells are cancer cells that use the telomerase enzyme
in their process of becoming immortal,36 (link) and telomerase is the target of our biosensor; thus, HEC-1-A represents
a positive control. To grow and subculture the HEC-1-A cell line,
we followed the manufacturer’s protocol37 and detailed it in the
Telomerase extract preparation was accomplished through the
following steps:38 (link) (1) cells were collected
in the exponential phase of growth, and 1 × 106 cells/mL
were washed twice with ice-cold phosphate-buffered saline (PBS) 1×
(140 mM NaCl, 2.7 M KCl, 10 mM NaHPO4, and 1.8 mM KH2PO4). (2) Then, cells were resuspended in ice-cold
3-[3-(cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS)
lysis buffer [10 mM Tris–HCl, pH 7.5, 1 mM MgCl2, 1 mM ethylenediaminetetraacetic acid (EDTA), 0.1 mM phenylmethanesulfonyl
fluoride (PMSF), 0.5% CHAPS, 10% glycerol] with a final concentration
of 25 × 106 cells/mL. (3) The suspension was then
incubated for 30 min on ice and centrifuged for 20 min at 15,000 rpm,
4 °C. (4) Finally, the supernatant was carefully transferred
to a fresh tube and frozen at −80 °C until use.