Seung Jae Jeong

RevTet-Off System Protocol2007. 6.20 Yu-Hong Min Guidelines for working with retrovirusesPut on double gloves.All waste must be autoclaved.Liquid waste : Do not suction. Collect in glass bottle then autoclave. PT67 packaging cell cultureDMEM, 10% tetracycline-free FBS, 1% P/G DoxycyclineDissolve at 1 mg/ml in water and filter sterilize. Store at…[Read more]

1)  On Frozen sectionsa. X-Gal Staining of Embryos frozen sections or Tibia Day 16.5/17.5 embryos were frozen on dry ice in tissue freezing medium (Leica) and cut into 15 µm sections. Tibiae were cut longitudinally. After fixation in 4% PFA for 5 min, specimen were incubated 24 hr in PBS containing 5 mM K3, 5 mM K4, 2 mM MgCl2, and 1mg/ml X-Gal…[Read more]

Bio Rad IEF cell(Model #PROTEAN IEF Cell) protocol                                                                                                                                                         Arthur   1. spin down the cell at 1,500Xg for 5 min2. wash the cell with ice cold PBS and lyse the cells with lysis buffer(1X PBS containing 1%…[Read more]

GST pull down assay A. Expression of GST fusion protein 1.     Inoculate 2ml of LB/antibiotics with 1 colony2.     O/N culture with shaking at 37℃3.     Inoculate pre-warmed 25ml of LB/antibiotics with all of 2ml-cultrure4.     1 hr at 37℃ with shaking vigorously5.     Inoculate pre-warmed 500 ml of LB/antibiotics with all of25ml-culture6.     Shak…[Read more]