Qproteome Mitochondria Isolation Kit

高純度ミトコンドリア調製物の効率的な分離用

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Qproteome Mitochondria Isolation Kit

Cat. No. / ID:  37612

12の高純度ミトコンドリア調製物用バッファーと試薬
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$540.00
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Qproteome Mitochondria Isolation Kitは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

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✓ Knowledgeable and professional Product & Technical Support

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Features

  • 高純度のインタクトなミトコンドリア
  • 標準化手順を使用する高い再現性
  • 簡便、迅速、使いやすいキットフォーマット

Product Details

Qproteome Mitochondria Isolation Kitキットは、標準的な卓上遠心分離機を使用する迅速で再現性の高い標準化された手順で、非常に高純度の完全活性ミトコンドリア調製物を提供します。

Performance

文献にある分画遠心分離法とは対照的に、細胞コンパートメントマーカータンパク質の分析により、Qproteome Mitochondria Isolation Kitでは、クロスコンタミネーションが実質的にないミトコンドリア画分が得られることが示されています(図「実質的にクロスコンタミネーションがないミトコンドリア画分」を参照)。2D-PAGE分析が示すように、細胞質基質汚染物質が実質的に存在しないため、存在量が少ない生物種の分析が非常に容易になり、穏やかな手順により、インタクトなミトコンドリアを含む高濃縮画分が得られます(図 「 インタクトなミトコンドリア膜を持つ高純度ミトコンドリア 」を参照)。完全性は、蛍光色素の取り込みを測定することによっても実証できます(図「1 mgタンパク質あたりより多くのインタクトなミトコンドリア」を参照)。
See figures

Principle

ミトコンドリアを扱う研究者が直面する大きな問題の一つとして、ミトコンドリア画分と細胞質基質画分の効率的な分離を挙げることができます。従来の調製法では面倒で時間がかかり、超遠心分離機を利用する必要があります。Qproteome Mitochondria Isolation Kitは、標準的なベンチトップ遠心分離機を用いた迅速で再現性の高い標準化された手順で、非常に高純度の完全活性ミトコンドリア調製物を提供します。

Applications

ミトコンドリアは、アポトーシス、プロテオミクス、生物医学研究、創薬など、数多くの分野で熱心に研究されています。Qproteome Mitochondria Isolation Kitは、そのタンパク質が本来の立体構造と完全な生物学的活性を保持している高純度ミトコンドリアを提供し、アポトーシス研究、2D-PAGE分析、その他の生化学的アッセイに非常に適しています。

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsSDS-PAGE、質量分析、活性アッセイ
Sample size5 x 10e6 ~ 2 x 10e7細胞、20 mg~60 mg
For glycoproteins: which type of glycoproteinsn.d
Binding capacity/yield20~80 µg
Species真核生物
Fractions isolated1分画
Start material細胞、組織

Resources

MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
クイックスタートプロトコール (1)
キットハンドブック (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