; d! Y2 \- _7 N, J, W7 I3 P您好！关于您的问题，我们咨询了总部的技术支持。其实我们自己在做茜素红染色的时候，也会有轻微的颜色差异。比如附件文件中的图7C和图10B，颜色也不是完全一样。但只要有阴性对照，实验结果应该还是OK的。" P$ \% {+ a$ }4 ^ B
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[attach]62633[/attach] $ e$ ^- p, C6 ]6 c& `2 ?/ `) w6 o0 s2 _/ U7 H6 I
此外您也可以参考下我们自己的茜素红染色操作步骤。谢谢！6 R9 R7 @8 q% p) j Y% w8 b8 T7 u
" Q. A* V4 ]. E% c+ N Alizarin Red S Staining Protocol:/ f5 U& l. T* a4 g# H
. W. u0 G0 m/ Z# P2 D/ zAlizarin Red S (Sigma A5533-25g)# x8 _( K1 r1 a$ g6 k' k* q! t
FW = 342.3g/mol ! m0 k; J, i5 M7 I D1 v6 F+ F- b7 X
Prepare 40mM Alizarin Red S Staining Solution (100mL):; J+ a9 Z( I9 j, C6 C! J% X: ?
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1. Dissolve 1.3692 g Alizarin Red S in 80mL of dH2O ; U* r! u8 U" \2. pH Alizarin Red S solution to 4.1 with 10% (v/v) ammonium hydroxide l2 E5 `1 G) H/ U
3. Top up volume to 100mL using dH2O * K. M' s* W' p1 l) R! F' f* ~ . |) B9 f6 a# X) d, A9 V! g" dStaining Protocol: (6 well plate) . g7 J0 R6 J" ?% ~, ^4 T % Q$ [9 G, ]6 m2 j& U! D1. Wash each well with 1xPBS) H: K3 {/ \# w- B
2. Fix cells using 10% formaldehyde at room temperature for 10min% _+ |# ?% y7 k
3. Wash cells two times with dH2O5 ]7 A) }; x: I+ d! H$ f: ]
4. Add 1mL of 40mM Alizarin Red S Staining Solution into each well and stain at room temperature for 20mins on a shaker $ w7 v# w( c1 I# F! b! M/ t5. Aspirate off excess stain and wash four times with 4ml of dH2O for 5mins each time on a shaker作者: liluoshushu 时间: 2014-8-20 12:54
回复 加拿大STEMCELL 的帖子 , m. n* ^$ y8 b! t v: e* `* }! m: P+ a
谢谢作者: kelvinlin 时间: 2018-1-29 11:29