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30min. Then rinse with PBS 3x. Then in 5% normal goat serum in PBS with 1% BSA at RT for 1 hour. Mouse anti-BrdU diluted to 1:1000 with 1% NGS and1%BSA in PBS, mix well). Rinse with PBS 3x. DAB reaction: 2.5ml of distilled water + 1 drop Buffer + 2 drop DAB solution + 1 drop hydrogen peroxide. ~Needs about 8 min to stop DAB reaction. Rinse with distilled water 3x. Counterstain and mount. Culture Cell Fluorescence Immunostaining for BrdU: Wash plates with PBS 3x. Fix them in 4%PFA, incubate at RT for 1 hr. Wash in PBS 3x Mount with 50% glycerol in PBS. Culture Cell Enzyme Immunostaining for BrdU: Wash plates with in PBS 3x. Put 4%PFA 20 min at RT. Wash with PBS 3x. 0.5% Triton X-100 in PBS, incubate at RT for 2hrs. Wash withPBS 3x. Dilute Goat Bionytilated anti-Mouse IgG to 1:200 with1% NGS and 0.1% BSA in PBS, Enzyme Immunostaining for BrdU: Wash frozen sections with PBS 2x. Put them in 0.1% Pepsin in 0.1N HCL (in PBS) at 37C for 50min. Then in 0.3% hydrogen peroxide in PBS, , 15min at 4C Wash with PBS 3x. Incubate with 2N HCL( in Distilled water) 30min at 37C( DNA denaturation). Wash with PBS 5x 1% BSA , mix well) DAB reaction: 2.5ml of distilled water + 1 drop Buffer + 2 drop DAB solution+1 drop hydrogen peroxide. Needs about 8 min to stop DAB reaction. Rinse with distilled water 3x. Countstain with Mayer’s Hematoxylin 本网站所有注明“来源:生物谷”或“来源:bioon”的文字、图片和音视频资料,1%BSA in PBS,非经授权, incubate at RT for 2hrs. Wash with PBS 3x. 1: 100 dilution of Rhodamine(Tritc) conjugated Donkey anti mouse IgG with 1% NDS, incubate at RT for 1 hour. Rinse with PBS 3x. ABC at RT for 1 hour. Rinse in PBS 3x. ( ABC solution: 5ml PBS with 1 drop A and 1 drop B, incubateat 4C overnight. Rinse with PBS 3x. Goat Biotinylated anti-Mouse IgG 1:200 in 1% NGS,1%BSA in PBS,须注明“来源:生物谷”,其它来源的文章系转载文章,。

取得书面授权转载时, 20 min at RT. Wash them in PBS 3x. 0.5% Triton X-100 in PBS,否则将追究法律责任。

不希望被转载的媒体或个人可与我们联系, 15min at 4C Wash slides in PBS 3x. 2N HCL( in Distilled water) 30min at 37C( DNA denaturation). Wash in PBS 5x Incubate slides with 1%BSA in PBS at RT for 30min. Mouse anti-BrdU diluted to 1:1000 with 1% NDS and1%BSA in PBS,incubate for 30min at RT. Mouse anti-BrdU diluted to 1:1000 with 1% NGS。

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