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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. / g' G) Z: B6 c2 I2 t) g/ y4 y4 U* |) P
Epithelial cells, endothelial cells, muscle cells, nerve cells, tumor cells : Z" l' r( O# U
7 B$ C6 Q0 R* d& L# P* DCaution:ECM gel may be stored up to 72 hours at 2-8 °C.! z* n! k- C1 A x6 Y0 ?
& G- b# [: G: e7 p2 a9 s5 ^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.
5 z/ |; h$ G5 p+ ?. Y. c
! s5 @7 [! l/ h% ?Properties6 u3 N: I6 ]7 z0 l
sterility dialyzed against chloroform
& a6 j7 }0 l$ m f# Zform liquid
. Q, n' q- z$ n. E+ d/ E! Z4 Uconcentration 8 - 12 mg/mL
5 l$ _- {" s# j% L5 L1 ~- psurface coverage 6‑10 μg/cm2
% b+ ^: ?# [3 {- K- ^total impurities endotoxin, tested # R& T! k& ]( O/ [
suitability cell culture tested $ b. [; G) U5 Z+ ~9 P3 B6 W6 P1 T0 g" `
shipped in dry ice
0 w- J$ v0 G d t# v; `, Qstorage temp. −20°C |
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