Cells were washed with 1X Phosphate-Buffered Saline (PBS; 21040-CV, Corning) once and then fixed with 10% neutral buffered formalin (89370–094, VWR) for 20 min at room temperature. After washing with 1X PBS, fixed cells were pre-incubated with 100% propylene glycol (P4347, Sigma-Aldrich) for 5 min and then stained with Oil Red O solution (I1516, Sigma-Aldrich) for 15 min. Then, cells were washed with 85% propylene glycol once and followed with 1X PBS twice. Cell images were taken using an inverted microscope. To quantify the amount of accumulated lipids, Oil Red O was extracted from the stained cells using 250 μl of 100% isopropanol and two aliquots of 100 μl were transferred to a 96-well assay plate (9017, Corning). The OD values were measured at 520 nm using a plate reader (Multiskan™ FC Microplate Photometer, Thermo Scientific).
P4347
Manufactured by Merck Group
P4347 is a multi-channel pipette designed for accurate and precise liquid handling. It features adjustable volume settings and is suitable for a wide range of applications in laboratory research and testing.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using p4347
1
Lipid Accumulation Quantification
2
Vapor Exposure System for Nicotine Studies
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CVE was performed using a vacuum-based vapor exposure system from La Jolla Alcohol Research17 (link),19 (link),20 (link). Animals were placed into sealed home cages with their standard housing cage-mates for 8 h a day, 5 days a week (9:00–17:00 M-F) before being returned to their standard home cages. The sealed cages continuously received clean room air from a down-stream vacuum pump (16 L/m), and a computer-controlled Electronic Nicotine Delivery System (ENDS) delivered vapor (3 s, 200 °C) at pre-defined inter-vape intervals (2, 5, 10, 15, and 60 min). The e-liquid vehicle (VEH) consisted of 50% propylene glycol (Sigma-Aldrich, P4347) and 50% vegetable glycerin (Sigma-Aldrich, G5516) (50/50 PGVG), and all experiments with nicotine (NIC) used a dose of 20 mg/ml (−)−nicotine (Sigma Aldrich, N3876). During all vapor exposure procedures, mice could move freely within the home cage with ad libitum access to food and water.
3
Oil Red O Staining of Zebrafish Larvae
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At 7 dpf, larvae were fixed with 4% paraformaldehyde in 1× PBS at 4°C overnight. After rinsing with 1× PBS 3 times, fixed larvae were incubated sequentially with 85% and 100% propylene glycol (P4347, Sigma-Aldrich) for 10 min at room temperature. Larvae were transferred to ORO staining solution [0.5% ORO (O0625, Sigma-Aldrich) in 100% propylene glycol] and placed on a platform rocker for overnight staining at room temperature. Stained larvae were destained with gradual transition from 100% propylene glycol to 1× PBS and eventually transferred to glycerol for imaging. Images were obtained using a Leica MZ16F stereomicroscope (Leica Microsystems) with an AxioCam HRc CCD camera (Carl Zeiss).
4
Vacuum-Based Vapor Exposure for Nicotine Studies
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CVE was performed using a vacuum-based vapor exposure system from La Jolla Alcohol Research17 (link),19 (link),20 (link). Animals were placed into sealed home cages with their standard housing cage-mates for 8 hours a day, 5 days a week (9:00–17:00 M-F) before being returned to their standard home cages. The sealed cages continuously received clean room air from a down-stream vacuum pump (16 L/m), and a computer-controlled Electronic Nicotine Delivery System (ENDS) delivered vapor (3 sec, 200°C) at pre-defined inter-vape intervals (2, 5, 10, 15, and 60 minutes). The e-liquid vehicle (VEH) consisted of 50% propylene glycol (Sigma-Aldrich, P4347) and 50% vegetable glycerin (Sigma-Aldrich, G5516) (50/50 PGVG), and all experiments with nicotine (NIC) used a dose of 20 mg/ml (−)-nicotine (Sigma Aldrich, N3876). During all vapor exposure procedures, mice could move freely within the home cage with ad libitum access to food and water.
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