Application：Recommended for use as a cell culture substratum. For a 24-well plate, use 230-250 μl/well. For a 96-well plate, use 50-100 μl/well. Thaw gel overnight at 2-8 °C before use. The thawed gel may be diluted up to two-fold with cold (2-8 °C) Dulbecco′s Modified Eagle′s Medium. Gel dilutions should be made before it is added to the plate. ECM will gel within 5 minutes at 20 °C. For prolonged manipulations, work should be conducted below 10 °C. Dispense gel to wells of a multiwell plate using pipettes pre-cooled to 2-8 °C. A gel forms at 37 °C and maintains this form with culture medium for at least 14 days. Cells may be plated on top of a thin gel layer (0.5 mm) or cultured inside a 1 mm layer. When cultured inside, cells should be added to the gel prior to plating at a recommended density of 3-4 × 104 cells per mL. To dissociate cells from the gel, use protease (dispase) dissolved in PBS without calcium, magnesium, and EDTA at a working concentration of 0.6-2.4 units/ml. + o" A( E7 h2 q u# ^2 u. Y4 b. b
Epithelial cells, endothelial cells, muscle cells, nerve cells, tumor cells / H; M1 t* a f; V7 r: J8 o
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Caution：ECM gel may be stored up to 72 hours at 2-8 °C. , Z$ }* u" f) U7 f5 A' W, Z$ F# d8 U" s! u% B) i0 u! i1 O
Other Notes：ECM gel is composed primarily of laminin, collagen type IV, heparan sulfate proteoglycan and entactin. Approximately 8-12 mg/ml basement membrane matrix protein in Dulbecco′s modified Eagle′s medium with 50 μg/ml gentamicin.& V; Z+ E& w/ F( I% i3 n( x. x
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Properties5 k- n1 F8 V5 O0 R/ q2 ~) Z
sterility dialyzed against chloroform 4 f K& `/ e7 a
form liquid 0 k% }& L; W) j5 T! m
concentration 8 - 12 mg/mL / v. D0 R5 q+ `7 W$ |surface coverage 6‑10 μg/cm2 ! q. u5 y* ?4 u; W( Y% n; g
total impurities endotoxin, tested ( [# y1 j' u; J7 H' N, ~# Osuitability cell culture tested ' B' w( j. @! [# xshipped in dry ice # V1 e# K8 `2 E E2 Z
storage temp. −20°C作者: hyx5415 时间: 2013-12-5 21:57