Cationic liposomes are traditionally used for the delivery of genetic materials such as various types of DNA (pDNA, cDNA, CpG DNA, oligonucleotide, antisense oligonucleotide, etc.), various types of RNA such as (siRNA, mRNA, etc.) and nucleic acid mimics (NAMs). The encapsulation of DNA into the conventional neutral charged PC based liposomes can be a technical problem mainly due to the plasmid size. Due to this problem in late 80s, the liposomes composed of cationic lipids and PE have been developed. The idea was to neutralize the negative charge of pDNA with positive charge of cationic lipids in order to capture more plasmid efficiently mainly due to electrostatic interaction and deliver them into the cells. Generally, the procedure is simply based on mixing the cationic liposomes with DNA or RNA and adding them to the cells. This results in the formulations of aggregates.
In order to design a proper cationic lipid for gene delivery, two approaches have been used for the cationic lipid synthesis: 1) cholesterol-based design such as DC-Cholesterol and GL-67 lipids, and 2) non-cholesterol-based designs such as DOTAB, DDAB and DOTMA. To successfully transfer the gene in vitro using liposomes, some consideration should be taken into account: i) the ability of binding and packing DNA/RNA in liposomes; ii) the interaction of the packaged DNA/RNA to the cell surface; iii) the efficiency of the internalization of DNA/RNA; iv) the intracellular DNA-release from endosomes in case of endocytosis involvement; v) the transgenic expression level in cell nuclei. pH-sensitive liposomes have been designed based on their tendency to release their content in the acidic condition. The primary concept was based on viruses that fuse with the endosomal membrane by means of a protein at pH 5-6, delivering their genetic material to the cytosol before reaching the lysosomes. Typically, a pH-sensitive liposome consists of dioleoylphosphatidylethanolamine (DOPE). Since phosphatidylethanolamine (PE) changes in acidic conditions, it is believed to act as a membrane fusion promoter. The effectiveness of the interaction between liposomes and cells is highly dependent on the liposome compositions. Liposomes are captured by various endocytosic processes, and the efficiency depends on the cell type and liposome size. Liposomes of various sizes and charges can attach to the macrophages and neutrophils through active phagocytosis. After attachment of the liposome to the cell surface, the internalization into the endosomes occurs due to a more acidic pH (6.50) at early endosomes. The liposomes are transferred to the last endosome with more acidic pH (5.5-6.0) by maturation or vesicular fusion, which takes 10-15 min. Twenty minutes (or more) after uptake, the contents are delivered to the lysosome with pH 5.0 or less. Lysosomes are the main degrading and last endocytotic section in the endocytotic pathway, in where pH-insensitive liposomes are accumulated and degraded. However, after penetration of pH-sensitive liposomes into cells, the accumulation and degradation do not occur.
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从1~2个T75培养皿的感染细胞培养液中纯化腺病毒
产品简介
本系列试剂盒可快速,高效地从腺病毒感染的细胞培养液中分离纯化腺病毒,相比于传统CsCl超速离心的腺病毒病毒纯化方式(需要24个小时才能完成纯化),本试剂盒可以在1个小时内完成病毒纯化,操作简单,快速,纯化率高,纯化到的病毒颗粒可直接用于下游实验,如细胞和动物感染。
产品特点
?操作简单,快速,可以在1个小时内完成病毒纯化,不依赖于超速离心操作。
?纯化效率高:小量纯化,可从1~2个T75瓶培养的细胞培养液中纯化病毒颗粒高达1x1012VPs
?每个纯化柱,可以重复利用一次,用于纯化相同种类的腺病毒。
保存条件
纯化柱和脱盐柱保存于4℃,其它组分室温保存。
试剂盒组分
Catalog#
V1160-01
Notes
Preps
10
MiniColumns
5
Canbeusedtwice
(Storeat4°C)
Press-OnCap
5
StoreatRT
DesaltingTube*
1
Canberegenarated
(Storeat4°C)
15mLCollectionTube
10
StoreatRT
10xWashBuffer
30mL
StoreatRT
2xElutionBuffer
30mL
StoreatRT
RegenerationBuffer
30mL
StoreatRT
联系方式:
Biomiga(中国)
电话:0573-82651206
QQ:441931287
联系人:张小姐
短截就是把枝条剪短,主要作用是促使其抽生新梢,增加分枝数目,以保证树势健状和正常结果。短截常用于骨干枝组修剪,结果枝组修剪,和树体局部更新复状。
短截按其长度可分为:
① 中短截:在一年生枝的中部短截,剪后萌发的顶端枝条,长势强,下部枝条长势弱。
② 重短截:剪去一年生枝的2/3。剪后萌发出的枝条较强状,一般用于主侧枝延长头修剪。
③ 重剪:剪去一年生枝的3/4-4/5,剪后萌发出的枝条长势强状,常用于发育枝作延长枝头和徒长果枝,中果枝的修剪。
④极重短截:剪去一年生枝的4/5以上,萌发后的枝条中庸偏状,常用于将发育枝和徒长枝培养结果枝组。
⑤留基部2芽剪:剪后萌发枝条较旺盛,常用于预备枝的修剪。对于幼龄树,树势较旺,以培养良好而牢固的树形结构,提早结果为主要目的,以轻短截,少疏间为主,从始果期到盛果期,主要使桃树多结果,并形成好的树形。
试剂里面含有的是病毒的抗原,现在基本都是人工合成或提取的,跟病毒完全是两回事
病毒液通过pre-filterdisc的时候是不是要制造一个真空环境产生压力差使液体通过?这个时候是用自备的针管还是试剂盒配的5毫升的针管啊?
它一般在医院、制药企业使用。
试剂盒使用示例: 试剂盒的产生正是为了使实验人员能够摆脱繁重的试剂配制及优化过程,所以试剂盒中一般配备有相应的使用说明书
过表达慢病毒
>10^8 PFU/ml
干扰慢病毒
>10^8 PFU/ml
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