网站标志
导航菜单
购物车
购物车 0 件商品 | 查看购物车 | 我的订单 | 我的积分 | 会员中心
当前日期时间
当前时间:
商品搜索
商品搜索:
价格
点评详情
点评详情
发布于:2018-7-4 00:40:09  访问:20 次 回复:0 篇
版主管理 | 推荐 | 删除 | 删除并扣分
Ll with the addition of 10 g/mL Sequa-brene (Sigma). After 2 h
Similarly, to characterize the purity of primary INK1117 price tonsil keratinocytes in culture, antibodies against CD3, CD4, CD11a/LFA1, CD32, CD64, CD89, and human fibroblast were used (Table 1). To detect the expression of -galactosidase, cells were stained with 1 mg/mL PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/27597769 X-Gal in 5 mM KFe4(CN6) 3H2O, 5 mM KFe3(CN6) 3H2O, and 1 mM MgCl2 and incubated at 37 for 2 h. A positive well contained two or more blue cells. Positive- and negativestained wells were tabulated and TCID50 was calculated using the Reed-Muench TCID50 calculation [68].Flow cytometry TERT-2 or TE cells were washed once with DPBS and incubated with 0.02 (W/V) EDTA for 10 min. Detached cells were washed twice with DPBS supplemented with 2 FBS (wash buffer), and resuspended at 5 to 10 ?105 cells in 200 L wash buffer. To identify putative HIV receptors and co-receptors, cells were incubated at 4 for 30 min with 1 g of anti-CD104, CXCR4, CCR5, galactosylceramide (GalCer), heparin sulfate (HSPGs), DC-SIGN, or macrophage mannose receptor (Table 1). Similarly, to characterize the purity of primary tonsil keratinocytes in culture, antibodies against CD3, CD4, CD11a/LFA1, CD32, CD64, CD89, and human fibroblast were used (Table 1). All antibodies were obtained from BD Pharmingen, except anti-GalCer (Chemicon), antiheparin sulfate (Seikagaku) and anti-human fibroblasts (Sigma). Cells were then washed twice with 1 mL wash buffer to remove unbound antibody. If needed, cells were PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/27597769 stained with goat anti-mouse IgG or IgM conjugated with fluorescein isothiocyanate (FITC) (Jackson ImmunoResearch Laboratories, West Grove, PA) in 200 L wash buffer at 4 for 30 min to detect primary antibodies. Isotype controls and other staining controls were included. After staining, cells were washed three times with 1 mL wash buffer, fixed in 200 L of 2 paraformaldehyde, and stored at 4 until analysis using a FACSVantage SE flow cytometer (BD Biosciences). HIV infection To infect with HIV-1, TERT-2 cells were plated in 96-well tissue culture plates (1.5 ?104 cells/well) and grown overnight in monolayers to 80?0 confluence and infected at a MOI 0.01 (TCID50 per seeded cells), for 0.5 to 120 h. Every 48 h, media were replaced with fresh growth media to maintain viability of TERT-2 and TE cells. In some experiments, viruses were heat-inactivated (HV) by incubating in a water bath at 70 for 3 h and used as a negative control. At indicated times, HIV-1 was aspirated. To remove surface-bound HIV-1, some cultures were treated with 0.05 trypsin/0.53 mM EDTA for 3 min at room temperature, and then an equal volume of soybeanPage 11 of(page number not for citation purposes)Retrovirology 2008, 5:http://www.retrovirology.com/content/5/1/trypsin inhibitor (250 g/mL; Invitrogen) in HBSS was added. Trypsinization did not appear to disrupt the monolayers, which were washed three times in HBSS and maintained in growth media. Some cells were sub-cultured for 3 to 8 passages post inoculation. In some experiments, cells were pre-treated with azidothymidine (AZT; 500 M; Sigma) for 2 h, or colchicine (500 M; Sigma) for 30 min and then inoculated with HIV-1. Colchicine was washed from cultures before HIV-1 was added, but AZT remained with TERT-2 cells during HIV incubation.
共0篇回复 每页10篇 页次:1/1
共0篇回复 每页10篇 页次:1/1
我要回复
回复内容
验 证 码
看不清?更换一张
匿名发表 
脚注信息
Copyright (C) 2015-2016 All Rights Reserved. 光明云南石斛商城管理系统 版权所有   滇ICP备11005439号-1
服务时间:周一至周日 09:00 — 18:00  全国订购及服务热线:0691-5161027 
联系地址:云南省西双版纳傣族自治州勐海县工业园区   邮政编码:666200