Murine 췌장 아일릿 격리

Protocol

Collagenase 준비 아래 표에 표시된 최종 농도를 만들기 위해 50 ML 원뿔 및 추가 절연 버퍼로 collagenase를 달다. vortexing하여 collagenase를 해산. (12 마우스 + 2 = 14 여분의 마우스 마우스 X 5 ML / 마우스 = 70 ML 총 collagenase 솔루션) (70 ML X 0.5 MG / ML 또는 collagenase P의 총 0.3 MG / ML = 35 MG 또는 21 MG) <table style="BORDER-RIGHT: #ddd 1px solid; BORDER-TOP: #ddd 1px solid; …

Materials

Material Name	Tipo	Company	Catalogue Number	Comment
Dithizone	reagent			Dissolve 5mg Dithizone in 5 ml of DMSO in a 50 ml conical tube. Add 45 ml of normal saline and mix gently. Filter with a 60 cc syringe and a 0.22 “m syringe filter into a new 50 ml tube. Store at 4 OC, until use. Add 1 ml to each 35 mm counting Grid-plate, when needed.
PBS	Buffer
HBSS	Buffer
Isolation soln.	Buffer			HBSS containing 10 mM of HEPES, 1 mM of MgCl2, 5 mM of Glucose, pH 7.4
Wash soln.	Buffer			Isolation buffer containing 1mM CaCl2
Collagenase P		Boehringer Mannheim-Roche	1213873
Dissection tools				Hemostat clamp (n=2), Forceps (n=2), Scissors (for skin incision an for taking out pancreas from adjacent tissue)
Syringe				sterile, 5ml, 1/mouse.
30 G needles				sterile, 1/syringe
siliconized glass vials w/Teflon cap		Fisher	213-018-54	sterile, 1/mouse
large sink strainer				sterile
1000 ml Beaker				sterile
Gauze pads				4×4, 2/mouse
plastic eye dropper				sterile
Mice	Animal