The Endoplasmic Reticulum Isolation Kit does not contain membrane proteins and peripheral membrane proteins. The kit includes the following components:
• Isotonic Extraction Buffer 5× (100 mL): Contains 50 mM HEPES (pH 7.8), 1.25 M sucrose, 5 mM EGTA, and 125 mM KCl.
• Hypotonic Extraction Buffer 10× (10 mL): Contains 100 mM HEPES (pH 7.8), 10 mM EGTA, and 250 mM KCl.
• Calcium Chloride Solution (5 mL): A 2.5 M CaCl2 solution.
• OptiPrep™ Density Gradient Medium (100 mL): A 60% (w/v) solution of iodixanol in water.
• Needle (4 inch, 20 gauge): One each.
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一般描述
内质网(ER)是由互连的小管、小泡和扁囊组成的网络。其在各种细胞特殊功能中发挥着重要作用，包括蛋白质合成、钙隔离、类固醇生产、糖原储备和生产、膜蛋白嵌入等。
应用
内质网分离试剂盒含有制备各种纯度的ER所需的所有试剂：粗面ER（微粒体）、Ca2+ 沉淀粗面ER (RER) 富集微粒体，以及密度梯度纯化所得的粗面ER (RER)和滑面ER (SER)。ER分离过程可采用细胞色素c还原酶检测试剂盒测定NADPH细胞色素c还原酶的活性进行监测（货号：CY0100）。这种酶是一种ER膜蛋白，常被用作ER标志物。分离所得的ER可用于研究P450系统和异源物代谢、脂质代谢，也可用于回收ER膜和腔蛋白。该试剂盒以采用大鼠肝脏、肾脏和脑，小鼠肝脏，兔肝脏、肾脏、脾、心脏和脑，以及Jurkat和HeLa细胞系进行过测试。
内质网分离试剂盒用于从哺乳动物组织或细胞培养物中有效富集功能性内质网（ER）。 分离出的ER可用于蛋白质合成、降解和代谢途径分析。 ER代谢异常包括发生于儿童的肉碱乙酰转移酶缺乏症、囊性纤维化和许多保留更多ER蛋白的病症。 适用于最多50 g的组织或约20 mL的包装组织。
特点和优势
- 专为研究规模应用配制的提取试剂 - 节省时间并减少浪费
- 包含氯化钙溶液 - 无需超速离心即可快速简单地沉淀粗糙ER
- 产生功能完整的细胞器 - 生成的 ER 适用于功能研究、脂质代谢和蛋白质分析
- 与用于结构确认的产品相兼容 - 与细胞色素 C 还原酶 (CY0100) 配套试剂盒测试一起轻松确认完整性
- 可用于从动物软组织和培养细胞中分离ER
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危险声明
危险分类
Eye Irrit. 2
储存分类代码
10 - Combustible liquids
WGK
WGK 1
历史批次信息供参考:
分析证书(COA)
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其他客户在看
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亚细胞组分离心分离是一种常用技术，广泛适用于多种类型的细胞和组织。由于不同细胞器的大小、形状和密度不同，因此通过离心，可从温和均质的样品中轻松分离和纯化细胞器组分。
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
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Does the Endoplasmic Reticulum (ER) obtained with the Endoplasmic Reticulum Isolation kit contain membrane proteins and peripheral membrane proteins?
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What level of purity of endoplasmic reticulum (ER) can be anticipated when utilizing the Endoplasmic Reticulum Isolation kit with low, medium, or ultra centrifugation?
1 answer-
When using low and medium-speed centrifugation steps, the resulting preparations are generally expected to be crude. To achieve an expectation of purity, it is advisable to follow the ultracentrifugal procedure. Notably, during quality control, no specific purity specifications were found. The Specification Sheet merely indicates that the kit must be suitable. Purity levels may fluctuate based on factors such as tissue type, specimen age, and other variables. If there are concerns about mitochondrial or other impurities, it would be prudent to test for mitochondrial markers in the collected ER fraction. Additionally, the level of purity is likely to vary depending on the carefulness with which the different fractions are collected.
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