The Bacterial Cell Lysis Kit has been developed for the extraction of biologically active, soluble proteins and inclusion bodies from bacterial cells. The Bacterial Lysis Buffer is a proprietary improvement on the lysozyme based lysis method, which allows extraction of soluble proteins and concurrent removal of nucleic acids (DNA & RNA) released during cell lysis. The kit’s lysis eliminates viscosity build-up, allowing effective clarification with lower centrifugal forces.
Bacterial Cell Lysis Buffer is based on organic buffering agents andutilizes a mild non-ionic detergent and a proprietary combination ofvarious salts and agents to enhance extraction and stability ofproteins. Depending on the application, additional agents such asreducing agents chelating agent, and protease inhibitors may be addedinto Bacterial Cell Lysis Buffer. This reagent has been tested for usewith several widely used bacteria including E. coli strains. Bacterial Cell Lysis Buffer eliminates the need for laborious mechanical lysis of bacterial cells and removal of DNA/RNA with nuclease treatments. The proprietary combination of this reagent provides a simple and versatile method of bacterial protein extraction and isolation of inclusion bodies.
The Bacterial Cell Lysis Kit (GB-176; 100, 250 or 500 preps) includes a separate tube of modified lysozyme preparation that contains nucleases and results in optimal lysis and minimal contamination. This kit contains all of the reagents needed to perform protein extraction.For extracting soluble proteins, one prep is suitable for extracting approximately 300 mg wet cell pellets for every 1 ml Bacterial Cell Lysis Buffer. When the kits are used for extracting soluble proteins from spheroplasts, one prep is suitable for extracting approximately 90 mg wet cell pellets for every 40 μl Bacterial Suspension Buffer. Bacterial Lysis Buffer is also available separately for further downstream applications.
Storage/Handling: Store at 4°C. Do NOT freeze. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.
GoldBio活体成像技术:早在1999年由美国哈佛大学Weissleder博士率先提出了分子影像学(molecularimaging,MI)的概念,即应用影像学的方法对活体状态下的生物过程进行细胞和分子水平的定性和定量研究。活体成像便是基于分子影像学孕育而生的,通过这个成像系统,可以观测活体动物体内肿瘤的生长及转移,感染性疾病的发展进程,特定基因的表达等生物学过程。活体成像技术主要采用生物发光(bioluminescence)与荧光(fluorescence)两种技术。★生物发光是用荧光素酶基因标记细胞或DNA。★荧光技术则采用荧光报告基团(GFP、RFP,Cyt及dyes等)进行标记。★这一技术对肿瘤微小转移灶的检测灵敏度极高,不涉及放射性物质和方法,非常安全。操作极其简单、所得结果直观、灵敏度高。
活体成像两种检测技术介绍活体成像特点优点缺点生物发光检测bioluminescence★荧光素酶(Luciferase)对基因、细胞和活体动物进行标记;★荧光素酶催化底物(例如荧光素钾盐)反应后,会产生化学发光。这种光是由化学反应而来,不需要激发光;★标记方法是通过克隆技术,将荧光素酶的基因插入到预期观察的细胞染色体内,通过对克隆细胞进行筛选,培养出能稳定表达荧光素酶的细胞株。再将细胞株转移至特定的小鼠体内形成模型。★特异性强,无自发荧光;★高灵敏度,在体内可检测到几百个细胞,检测的深度在3-100px;★定量精确 ★信号较弱,检测时间较长;★仪器精密度要求较高;★细胞或基因需要转基因标记;★不可用于人体,不适用于抗体、多肽等标记荧光检测fluorescence★采用荧光报告基因(GFP、RFP等)或荧光染料进行标记;★需要外接激发光源,利用报告基因、荧光蛋白质或染料产生的荧光,就可以形成体内的生物光源。★荧光染料、蛋白标记能力强;★信号强,成像速度快,操作简便,实验成本较低;★未来可用于人;★适用范围广,可以是动物、细胞、微生物,也可以是抗体、药物、纳米材料等。★存在自发荧光,影响灵敏度;★光容易被动物组织吸收;★检测深度受限;★背景光干扰,定量准确度低
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根据组成不同,可分为两种,弱酸及其对应的强碱弱酸盐,弱碱及其对应的强酸弱碱盐。
因为HF可以和NaOH反应生成NaF和水,当NaOH反应完之后,NaF就可以与HF组成缓冲溶液,所以说可以直接使用NaOH和HF来配制缓冲溶液。
这也是一般配制缓冲溶液的方法,也就是用强碱和弱酸(或者强酸和弱碱)来配制缓冲溶液。
(2)酸和盐浓度等比例也增减时,溶液的pH值不便。
(3)酸和盐浓度相等时,缓冲液的缓冲效率为最高,比例相差越大,缓冲效率越低,一般地说缓冲液有效缓冲范围为PK±1pH
浙江大学奚振宇、徐又一、朱利平等2009年在journal of membrane science(中文翻译为《膜科学杂志》)发表的一篇论文,其中探讨了缓冲溶液的pH值对聚乙烯-多巴胺复合膜亲水性的影响,发现缓冲溶液的pH值为8.5时制备的复合膜亲水性最强。
通过大量的实验,研究人员发现,pH值为8.5时制备的多巴胺复合膜性能最优。这个是通过大量的实验总结得到的结论。
2.尿素:有膜蛋白的损失
3.RIPA:用于磷酸化蛋白的温和裂解,不破坏磷酸基团