Calcium Flux Assays using the LabChip 3000
...n 1.5 g/L sodium bicarbonate with 100 U/mL penicillin, 0.1 mg/mL streptomycin, 2 mM L-glutamine, 0.1 mg/mL Geneticin G418, 1 mM sodium pyruvate, 10 mM hepes
and 10% Fetal Bovine Serum.
Cells were maintained at 37 oC and 5% CO 2 and subcultured in monolayers in T75 flasks using standard cell cult...
Monitoring Receptor-Mediated Changes in [Ca2+]i Using Single-Wavelength and Dual-Wavelength Indicators on the FlexStation Workstation
...Salt Solution (HBSS) (10X) (Invitrogen Life Technologies, cat # 14065-049), hepes
1M buffer solution (Invitrogen Life Technologies, cat # 15630-056), FBS: (F...ash buffer: this buffer contains a final concentration of 1X HBSS and 20 mM hepes
diluted in sterile d.H 2 O. Important: To prevent dye extrusion by the anio...
Gq-coupled receptor assays using FLEXstation: M1 muscarinic receptor
... the microplate were treated with
0.5 M carbachol, while cells in the other half of
the plate were treated with 1X Reagent Buffer
(1X HBSS/20 mM hepes
pH 7.4). The
FLEXstation Calcium Assay Kit was used to
monitor carbachol-mediated calcium
mobilization (Figure 5). Z factor was calculated
Gq-coupled Receptor Assays Using FLEXstation
...lution (Gibco/Invitrogen, cat# 25200-072; Tel: 1800-828-6686), Hank's balanced salt solution (HBSS) (10X) (Gibco, cat# 14065056; Tel: 1-800-828-6686), hepes
1M buffer solution (Gibco, cat#15630-080).
CF2Th cells were grown in D...
FLIPR Calcium 3 Assay Kit - An Optimal No-Wash Fluorescence Calcium Indicator Reagent for Non-Optimal Calcium Responses
...he FLIPR system. Growth medium was removed by aspiration and cells were washed twice with 150 l Hanks buffered saline solution supplemented with 20 mM hepes
and 2.5 mM probenecid (HBSS). Dye loading buffer comprised FLUO-4-AM at 2 M final concentration, 1% fetal bovine serum and 0.1% pluronic acid. After w...
ATP Quantitation in the LMax Microplate Luminometer (MaxLine Application Note #40)
...TM ATP Assay System, Cat. No. FF2000; Promega Corporation. 608277-2516. The kit contains Luciferase/Luciferin (L/L) reagent, ATP standard 10 -7 M in hepes
buffer, pH 7.75) and ATP-free water.
3. Solid white 96-well microplates; e.g. Porvair PS white, E&G Wallac Cat. # 204003; Tel: 1-800-221-9650; ...
Ca++-Induced G Protein Coupled Receptor (GPCR) Activation Monitored via Aequorin Luminescence in the LMax Microplate Luminometer (MaxLine Application Note #42)
...0, Glutamine Pen-Strep Solution Cat. No. 9316, G418 sulfate #98529, and 1 M hepes
buffer solution Cat. # 9319 were obtained from Irvine Scientific Tel. 1-800...s (2.5 M) were prepared by dilution in Hanks Balanced salt solution + 20 mM hepes
(H&H) with 1 mg/ml BSA.
PREPARATION OF CELLS
measurement of green fluorescent proteins and a red fluorescent protein in live cells in the Analyst GT multimode reader
...Buffer (1X Hanks Balanced Salt Solution with 20 mM hepes
buffer): Made from 10X Hanks Balanced Salt Solution (100 mL), 1M hepes
(20 mL) and cell culture treated water (880 mL)
...Gibco/ Invitrogen Cat. #14065-056)
(Irvine Scientific Cat. #9319)
FREE FAT CELL ASSAY
...lyse to remove ions. Dilute to 500 mL.
pH 7.9: 11.92 g hepes
is dissolved in 50 mL water. Adjust pH to 7.9 using 1N NaOH. Dilute to 100 mL. hepes
Buffer: 40 mL Krebs, 50 mL 10% HSA, 10 mL 0.5M...
Use of FlashPlate Technology for In Vitro Measurement of [35S]-GTPγS Binding in CHO Cells Expressing the Human 5-HT1B Receptor
... cells were pre-incubated at 30C for 30 minutes in hepes
[20 mM], pH 7.4, in the presence of MgCl2 [3 mM], ... filtration, followed by five washes with ice-cold hepes
buffer. Bound radioactivity was determined by liqu... a desired membrane protein concentration in 25 mM hepes
buffer containing 2.5 mM CaCl2 and 1 mM MgCl2, at ...