Qproteome Mitochondria Isolation Kit

用于高效分离高纯度线粒体制剂

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Qproteome Mitochondria Isolation Kit

Cat. No. / ID:  37612

用于 12 种高纯度线粒体制剂的缓冲液和试剂
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Qproteome Mitochondria Isolation Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

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✓ Knowledgeable and professional Product & Technical Support

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Features

  • 完好的高纯度线粒体
  • 采用标准化程序,可重现性高
  • 方便、快捷且易用的试剂盒形式

Product Details

Qproteome Mitochondria Isolation Kit 按照快速、可重现的标准化程序,使用一台标准台式离心机提供纯度极高的完全活性线粒体制备。

Performance

与文献中的差速离心法相比,对细胞区室标记蛋白的分析表明,Qproteome Mitochondria Isolation Kit 提供的线粒体馏分几乎没有交叉污染(见图“线粒体馏分几乎没有交叉污染”)。如 2D-PAGE 分析所示,由于几乎不存在细胞质污染物,因此大大方便了对低丰度物种的分析,而且这种温和的程序可提供含有完好线粒体的高富集馏分(见图“ 具有完整线粒体膜的高纯度线粒体 ”)。还可以通过测量荧光染料的吸收来证明完整性(见图“每毫克蛋白中含有更多完好的线粒体”)。
See figures

Principle

线粒体研究人员面临的一个主要问题是,如何高效地分离线粒体和细胞质部分。传统的制备方法繁琐、耗时,而且需要使用超速离心机。Qproteome Mitochondria Isolation Kit 按照快速、可重现的标准化程序,使用一台标准台式离心机提供纯度极高的完全活性线粒体制备。

Applications

线粒体是细胞凋亡、蛋白质组学、生物医学研究和药物发现等多个领域的热门研究课题。Qproteome Mitochondria Isolation Kit 可提供高纯度线粒体,其蛋白保留了原生构象和完全生物活性,因此非常适合凋亡研究、2D-PAGE 分析和其他生化检测。

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsSDS-PAGE、质谱法、活性检测
Sample size5 x 10e6 至 2 x 10e7 个细胞,20 mg - 60 mg
For glycoproteins: which type of glycoproteinsn.d
Binding capacity/yield20-80 µg
Species真核细胞
Fractions isolated一个馏分
Start material细胞、组织

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (1)
For purification of mitochondria from eukaryotic cells and tissues
Safety Data Sheets (1)
Certificates of Analysis (1)
Kit Handbooks (1)
For purification of mitochondria from eukaryotic cells and tissues

FAQ

Can mitochondria separated with the Qproteome Mitochondria Isolation Kit be used for downstream DNA isolation?

Even though we have not tested this, we assume that both the QIAamp DNA Micro Kit and the AllPrep DNA/RNA Mini Kit will work to isolate DNA from mitochondria separated with the Qproteome Mitochondria Isolation Kit.

 

FAQ ID -1188
Which Qproteome or Protein Fractionation Kits from QIAGEN are compatible with tissue samples?

The Qproteome Cell Compartment Kit, the Qproteome Nuclear Protein Kit, the Qproteome Mitochondria Isolation Kit, and the PhosphoProtein Purification Kit are compatible with tissue samples. The respective protocol can be found in the respective handbook.

The Qproteome Mammalian Protein Prep Kit is also compatible with tissue, there is the QIAGEN Supplementary Protocol: Purification of protein from animal tissues using the Qproteome Mammalian Protein Prep Kit and the TissueRuptor™ available. For all these protocols the TissueRuptor is used.

 

 

 

FAQ ID -755
Can the nuclei and cell debris fraction generated in the QProteome Mitochondria isolation procedure be used for further fractionation?

We have not tested this application. We would suggest to split the sample, and perform different fractionation procedures separately on the two sample portions.

If splitting the sample is not feasible, then it should theoretically be possible to use the pellet generated at step 7 of the QProteome Mitochondria Isolation Kit protocol (containing cell debris, nuclei and unbroken cells) as starting material for the Qproteome Nuclear Protein Kit.

Please note that this application will have to be tested and optimized by the enduser.

 

FAQ ID -1185
Is it possible to collect intact mitochondria before or after the density gradient centrifugation step of the Qproteome Mitochondria Isolation protocol?

Highly pure and intact mitochondria can be collected after the density gradient centrifugation at step 11b of the Qproteome Mitochondria Isolation protocol.

The fraction collected at step 9 of the protocol, prior to the cushion centrifugation, contains intact as well as broken mitochondria, and may also comprise structures of the ER, Golgi and plasma membranes.

FAQ ID -1187
How many cells can be used with the Qproteome Mitochondria Isolation Kit, and how much mitochondrial protein can be expected?

The Qproteome Mitochondria Isolation Kit can be used with 5 x 106 to 2 x 107 cells. Depending on the cell line, the yield from a single fractionation procedure is 20–80 µg protein from 5 x 106 cells.

Protein can be isolated from the mitochondria using, e.g. the QProteome Mammalian Protein Prep Kit, or home-made lysis buffers.

 

 

FAQ ID -1189
Do you have a kit for protein isolation from mitochondria prepared with your Qproteome Mitochondria Isolation Kit?

Yes, you can use the Qproteome Mammalian Protein Prep Kit to isolate proteins from mitochondria separated with the Qproteome Mitochondria Isolation Kit.

 

FAQ ID -1190
Do I need an ultracentrifuge for the density gradient centrifugation step in the Qproteome Mitochondria Isolation protocol?
No, an ultracentrifuge is not required for the Qproteome Mitochondria Isolation Kit. All centrifugation steps are performed in a standard table-top centrifuge.
FAQ ID -1186
What starting material can be used with the Qproteome Mitochondria Isolation Kit?

The Qproteome Mitochondria Isolation Kit has been developed for cultured mammalian cells. We have tested HeLa, Cos7, NIH, HEK 293 and SH-SY5Y cell lines.

We have not tested animal tissue, yeast, or plant cells. Animal tissue might work if a method for cell disruption that leaves the mitochondria intact can be found. Unfortunately, we do not have any protocol recommendations for these starting materials.

FAQ ID -1184