| 查看: 7138 | 回复: 8 | ||
sanger09新虫 (小有名气)
|
[求助]
求细胞粘附实验的具体步骤
|
|
各位大神，我本科不是学生物的，跨专业考过来的，现在要做个细胞粘附实验。看了好久，不知道怎么做，所以来小木虫发帖询问。 各位能够告诉我，细胞粘附实验的具体步骤是怎么样的？ 越详细越好。 |
回复此楼» 猜你喜欢
售SCI-T0P文章，我:8O.5.5.1.O.54,科目齐全,可+急
已经有3人回复
-
售SCI文章，我:8O5.5.1.O.54,科目齐全,可+急
已经有3人回复
-
售SCI文章，我:8O.5.5.1O.54,科目全,可十急
已经有3人回复
-
售SCI一区T0P文章，我:8O.55.1.O.5.4,科目齐全,可+急
已经有3人回复
-
售SCI一区T0P文章，我:8.O.55.1.O.5.4,科目全,可+急
已经有3人回复
-
售SCI文章，我:8O.5.5.1O.54,科目全,可十急
已经有3人回复
-
售SCI文章，我:8O5.5.1.O.54,科目齐全,可+急
已经有3人回复
-
售SCI一区文章，我:8O5.5.1.O5.4,科目全,可伽急
已经有3人回复
-
售SCI-T0P文章，我:8O.5.5.1.O.54,科目齐全,可+急
已经有3人回复
-
售SCI一区T0P文章，我:8.O55.1.O.54,科目全,可十急
已经有4人回复
» 本主题相关价值贴推荐，对您同样有帮助:
- 最新Adv. Funct. Mater上的一篇有意思的报道：细胞和微生物的差异性粘附
已经有12人回复
- 生物降解生物相容的弹性体（细胞粘附不好）的材料可以做什么材料呢？
已经有3人回复
- 如何让PC12细胞粘附在PEG水凝胶表面
已经有3人回复
- 【求助】血小板粘附实验的问题
已经有6人回复
- 【推荐】973 三维培养干细胞自我更新与定向分化的调控网络
已经有9人回复
- 【求助】无毒性，细胞粘附却不好，这种材料能做什么，或者如何改进？
已经有5人回复
- 【求助】怎样对材料上的细胞做荧光染色
已经有7人回复
- 微生物学考研资料1(沈萍版同步）免币分享
已经有67人回复
sanger09
新虫 (小有名气)
- 应助: 0 (幼儿园)
- 金币: 16.4
- 散金: 71
- 帖子: 183
- 在线: 41.3小时
- 虫号: 1062371
- 注册: 2010-07-22
- 专业: 功能与智能高分子
2楼2012-06-03 21:21:09
peopole
银虫 (小有名气)
- 应助: 2 (幼儿园)
- 金币: 1000.2
- 散金: 50
- 红花: 2
- 帖子: 231
- 在线: 72.4小时
- 虫号: 1211637
- 注册: 2011-02-24
- 性别: GG
- 专业: 水产基础生物学
3楼2012-06-04 08:56:53
xuzhengxin405
至尊木虫 (著名写手)
- BioEPI: 2
- 应助: 5 (幼儿园)
- 金币: 18844.6
- 帖子: 1068
- 在线: 324.7小时
- 虫号: 153867
- 注册: 2006-01-03
- 性别: GG
- 专业: 药学
【答案】应助回帖
★ ★ ★
感谢参与，应助指数 +1
zhaohq1209: 金币+3, 有帮助~学习了~ 2012-06-05 08:40:35
感谢参与，应助指数 +1
zhaohq1209: 金币+3, 有帮助~学习了~ 2012-06-05 08:40:35
|
给你两个protocols，我当时就是参照这些资料做的 Adherence assay CHOK1 cells were grown in 24-well tissue culture plates to approximately 80% confluence. Cells were transfected with either pCMV5 Nrx1β-mCherry or pCMV5 FLAG-Nrx1β-mCherry using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions and grown for 36 hours to allow for protein expression. For attachment assays, bacteria were washed 2 times in PBS, resuspended in CD CHO media and incubated for 1 h with CHOK1 cells at an MOI of 10. To remove unattached bacteria, cells were washed 4 times with PBS. To determine the number of attached bacteria, CHOK1 cells were lysed with sterile water and serial dilutions plated on the appropriate media. The number of attached bacteria was reported as a percent of total bacteria at the end of the incubation period. Each experiment was performed in triplicate wells and repeated three times. Statistical analysis was performed using the Student's t-test. Adhesion assays A549 cells (human alveolar type II pneumocytes; ATCC CRL-185), were grown to 90% confluence on 24-well plates (~106 cells/well). Prior to use, cells were washed with cell F12 media to remove serum. For competitive inhibition binding assays, A549 cells were incubated with 1?M of either rBR, rBR.C, a synthesized peptide corresponding to AA 122-167, or BSA for 1 HR. Following incubation, cells were exposed to media that contained 107 cfu/mL of bacteria and incubated for 1 h at 37°C in 5% CO2. Nonadhering bacteria were removed by washing the cells 3 times with T-PBS and the number of adhering bacteria was determined by lysis of the monolayer with 0.1% Triton X-100 and plating wells per experiment. |
4楼2012-06-04 09:35:29
castlered
木虫 (小有名气)
- 应助: 9 (幼儿园)
- 金币: 1461.7
- 帖子: 245
- 在线: 306.5小时
- 虫号: 460224
- 注册: 2007-11-15
- 性别: MM
- 专业: 病原细菌与放线菌生物学
5楼2012-06-04 14:51:43
xuzhengxin405
至尊木虫 (著名写手)
- BioEPI: 2
- 应助: 5 (幼儿园)
- 金币: 18844.6
- 帖子: 1068
- 在线: 324.7小时
- 虫号: 153867
- 注册: 2006-01-03
- 性别: GG
- 专业: 药学
6楼2012-06-04 23:07:15
7楼2012-06-20 11:10:41
8楼2013-03-16 16:42:31
9楼2013-03-16 16:44:14
20