http://www.paper.edu.cn
(kindly provided by Prof. Chris Wright in Vanderbilt University,USA) was amplified
by PCR. Briefly, PCR was performed with the following degenerated primers
including an upper primer 5’- GGGGATCCATGAACAGTGAGGAGCAGTAC-3’
and a lower primer 5’- GCTCTAGATCACCGGGTTCCTGCGG-3’. The amplified
cDNA was subcloned into pCDNA3.1(+). The construct was transfected into cells
using Lipotectamine-2000 (Invitrogen,
Carlsbad, CA). Cells were cultured for
additional 48h.
In Situ Hybridization. The oligonucleotide antisense probes used for in situ studies
were 5'-CCCAGAGGGCAAGCAGGGCCAGCAGGGGTAGGA and
5'-TCACGGCGGGACTTGGGTGTGTAGAAGAAGCC. In situ hybridization was
carried out using 2 pmol/ml labeled oligonucleotides for
18 h at 40°C. Bound probe
was visualized using Texas Red Tyramide
Signal Amplification detection reagent
(Boster Biological Technology Ltd, China)
according to the manufacturer's
instructions. The cells were counterstained with hematein to visualize nuclei.
Immunocytochemistry. Cells were washed with
0.01 mol/l PBS three times for 3 min
and fixed by 4% araformaldehyde. The specific
binding was inhibited by 5% goat
serum in 0.01 mol/l
PBS at room temperature for 60 min in a humid chamber. A rabbit
insulin antibody against the insulin in the C2C12
(1:100 diluted with 0.1% Triton
X-100 in 0.01 mol/l PBS) was added to slides and incubated at 4°C overnight
in a
humid chamber. After washing, cells were incubated with
a biotin-conjugated goat
-5-
剩余20页未读，继续阅读
weixin_38609732
- 粉丝: 8
- 资源: 964
我的内容管理
展开
最新资源
- Vue实现iOS原生Picker组件：详细解析与实现思路
- Arduino蓝牙小车：参数调试与功能控制
- 百度Java面试精华：200页精选资源涵盖核心知识点
- Swift使用CoreData填坑指南：CoreData在Swift 3.0的变化
- 微距离无线充电器创新设计及其实验探索
- MTK Android平台开发全攻略：44步详解流程
- RecyclerView全面解析：替代ListView的新选择
- Android开发：自动适配中英文键盘解决方案
- Android调用WebService接口教程
- Android开发：BitmapUtil图片处理全解析与实例
- Android多线程断点续传实现详解
- PCA算法在人脸识别会议签到系统中的应用
- EventBus 3.0：Android事件总线详解与实战应用
- Android FileUtil：全面解析文件操作实用技巧与实例
- RecyclerView添加头部和尾部实战教程
- Android实现微博滑动固定顶部栏实战与优化
资源上传下载、课程学习等过程中有任何疑问或建议，欢迎提出宝贵意见哦~我们会及时处理！
点击此处反馈
