Table of Materials
| Name of Material/Equipment | Company | Catalog Number |
Comments/Description |
|---|---|---|---|
| Coating Solution: | |||
| DMEM | Gibco | 10569010 | Always add gentamicin (1:1000 by volume) prior to use; 24 mL |
| HAMS F12 | Lonza | 12-615F | Always add gentamicin (1:1000 by volume) prior to use; 24 mL |
| Collagen | Life Technologies | A1064401 | 1.7 mL |
| Matrigel | Fisher | CB40234A | 1 mL |
| Plating Media: | |||
| DMEM | Gibco | 10569010 | Always add gentamicin (1:1000 by volume) prior to use; 12.5 mL |
| HAMS F12 | Lonza | 12-615F | Always add gentamicin (1:1000 by volume) prior to use; 12.5 mL |
| Heat Inactivated FBS | Life Technologies | 16000044 | 20 mL; can be purchased as regular FBS and heat-inactivated by placing in a 40 °C water bath for 20 minutes |
| Amniomax | Life Technologies | 12556023 | 5 mL |
| Myoblast Media: | |||
| DMEM | Gibco | 10569010 | Always add gentamicin (1:1000 by volume) prior to use; 17.5 mL |
| HAMS F12 | Lonza | 12-615F | Always add gentamicin (1:1000 by volume) prior to use; 17.5 mL |
| Heat Inactivated FBS | Life Technologies | 16000044 | 10 mL; can be purchased as regular FBS and heat-inactivated by placing in a 40 °C water bath for 20 minutes |
| Amniomax | Life Technologies | 12556023 | 5 mL |
| Differentiation Media: | |||
| DMEM | Gibco | 10569010 | Always add gentamicin (1:1000 by volume) prior to use; 24 mL |
| HAMS F12 | Lonza | 12-615F | Always add gentamicin (1:1000 by volume) prior to use; 24 mL |
| Heat Inactivated Horse Serum | Sigma | H1138 | 1.5 mL |
| Insulin-Selenium-Transferrin | Life Technologies | 41400045 | 0.5 mL |
| Other Materials: | |||
| PBS | Gibco | 14040133 | |
| Gentamicin | Sigma | G1397 | |
| TrypLE | Gibco | 12604013 | |
| DMSO | Sigma | 472301 | Prepare as 10% DMSO in Myoblast Media for freezing cells |
| Forceps | Any | ||
| Razor Blades | Any | ||
| Scissors | Any | ||
| Whatman paper | VWR | 21427-648 | |
| 60 mm plate | VWR | 734-2318 | |
| 10 cm plate | VWR | 25382-428 (CS) | |
| T25 Flasks | ThermoFisher | 156367 | |
| T75 Flasks | ThermoFisher | 156499 | |
| Centrifuge Tubes (15mL) | BioPioneer | CNT-15 | |
| Oxygen Consumption Rates: | |||
| Seahorse XFe96 Analyzer | Agilent | Seahorse XFe96 Analyzer | Instrument used to measure oxygen consumption rates read out by acidification of the extracellular media |
| Seahorse XFe96 FluxPak | Agilent | 102416-100 | 96-well plates for use in XFe96 Analyzer |
| Seahorse XF Cell Mito Stress Test Kit | Agilent | 103015-100 | components may be purchased from other suppliers once assay is established; some recommendations are listed below |
| Seahorse XF Palmitate-BSA FAO substrate | Agilent | 102720-100 | components may be purchased from other suppliers once assay is established; some recommendations are listed below |
| Palmitic acid | Sigma | P5585-10G | for measurement of fatty acid oxidation |
| carnitine | Sigma | C0283-5G | for measurement of fatty acid oxidation |
| Etomoxir | Sigma | E1905 | for measurement of fatty acid oxidation |
| BSA | Sigma | A7030 | used as control or in conjugation with palmitic acid for use in measurement of fatty acid oxidation |