本帖最后由 加拿大STEMCELL 于 2014-7-8 16:21 编辑 & I5 c5 @8 A1 _4 u ; T, a2 e! y. I# }2 [3 Q! G ]& q3 T回复 liluoshushu 的帖子 9 V4 S9 x# O% _( E* k6 ]0 ^$ {1 a3 f7 B0 @4 Y; F' q
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。 9 C! u. ^- \: ?$ w, B2 H8 n; E . I7 p2 ? e0 g[attach]62633[/attach]: i& v1 }2 {! q2 z/ e9 _
' b6 I7 r% Q" Z+ D W
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！ . X# o( n3 o4 O6 b! \4 \# ~( y. z% g E) X6 |) a Alizarin Red S Staining Protocol: ! s: D; \: g! Q) Z, |$ ? " k. c; X4 P3 F* m, n& F w5 O9 rAlizarin Red S (Sigma A5533-25g) , y* W' a9 c; oFW = 342.3g/mol) x8 m2 |2 y+ A) Z: P/ u
3 I7 W/ m* q% o8 WPrepare 40mM Alizarin Red S Staining Solution (100mL): ; b. g" W) Z( i- r ' z; `2 C+ s) i1. Dissolve 1.3692 g Alizarin Red S in 80mL of dH2O 6 e7 c& P# s& R$ T9 J2. pH Alizarin Red S solution to 4.1 with 10% (v/v) ammonium hydroxide4 x$ Q/ r8 ]: s0 J
3. Top up volume to 100mL using dH2O7 f! x( c( G/ j" y0 c9 ~
7 n" A2 P1 R y% J
Staining Protocol: (6 well plate)" t/ d) Q0 n: X" M& r/ K- l, f
$ P7 @9 I5 n# s; N" k1. Wash each well with 1xPBS# M& ~$ Y& l. Q! X/ R/ J
2. Fix cells using 10% formaldehyde at room temperature for 10min " j& i' l' }* L) ]) l9 j3. Wash cells two times with dH2O) S( c5 t* |6 }% z: m7 r
4. Add 1mL of 40mM Alizarin Red S Staining Solution into each well and stain at room temperature for 20mins on a shaker , i8 T1 l0 a& `( i5. 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