Microcentrifuge tubes

C57BL/6J mice were purchased from the Jackson Laboratory (Bar Harbor, ME) and housed in the Maine Medical Center Research Institute (MMCRI) AAALAC accredited animal facility. Bone marrow cells (BMCs) were collected from 8- to 10-weeks old female or male mice, and pooled for testing. Briefly, mice were euthanized by isoflurane over-dose followed by cervical dislocation. Femurs and tibias were isolated and the ends were trimmed off. BMCs were collected by centrifugation of bones at 13,000rpm for 30 s in 1.5 ml microcentrifuge tubes (USA Scientific), followed by depletion of red blood cell (RBC) with RBC lysis buffer (Sigma-Aldrich, MO). Cells were cultured in α-MEM (Corning) containing 1× GlutaMax (Gibco), 10% of fetal bovine serum (FBS) (Atlantic Biologics) and 1× antimycotic-antibiotic mixture (Gibco). Fourteen to eighteen hours later, non-adherent cells were collected and replated onto 10cm tissue culture dishes. The first overnight adherent cells were replenished with fresh medium. Three days later, adherent cells (1adBMC from overnight culture, and 2adBMC from overnight non-adherent cells) were collected using trypsin or Accutase (Millipore), and counted for OC differentiation, FACS analysis, or magnetic beads cell purification procedures. The Institutional Animal Care and Use Committee of MMCRI approved all protocols involving animals.

Formic Acid, Ammonium Hydroxide (30%), Propranolol Hydrochloride, Tamoxifen, 4 HydroxyTamoxifen and the UV Photochemical Reactor Enhanced Detection tube unit (PHRED) were obtained from Sigma Aldrich, St. Louis MO, USA. The HPLC grade reagents, methanol, acetonitrile, deionized water, Bovine serum albumin (BSA), Potassium Phosphate, 12×75mm polystyrene tubes from Fisher Scientific, Pittsburg, PA, USA. Endoxifen and ND-Tamoxifen, were obtained from Toronto Research Chemicals, Ontario, Canada. The Solid Phase Extraction columns (SPE), STRATA-X-C 3, 3u, were obtained from Phenomenex, Torrance, CA, USA. The SPE Extraction Manifold was obtained from Varian, Walnut Creek, CA, USA. The HPLC Column, and guard cartridge, Spherisorb C18 CNRP, 4.6×250mm, 5u, from Waters, Milford, MA, USA. The Micro centrifuge tubes were obtained from USA Scientific, Ocala, FL, USA.