Qproteome Cell Compartment Kit

細胞の位置に応じたタンパク質の分画用

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Qproteome Cell Compartment Kit

Cat. No. / ID:  37502

For up to 10 subcellular fractionations*: Extraction buffers, Protease Inhibitor Solution, Benzonase®.
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¥50,500
Qproteome Cell Compartment 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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Product Details

真核細胞は複雑で、整然とした、高度に構造化されたシステムです。Cell Compartment Kitは、インタクトな真核細胞や組織を迅速かつ容易に細胞内分画できるように設計されています。細胞ペレットにさまざまな抽出バッファーを連続添加することにより、さまざまな細胞コンパートメント内のタンパク質を選択的に分離できます。

Performance

Qproteome Cell Compartment Kitを使用すると、分離した細胞(図「 マーカータンパク質の特異的分離」および「 さまざまな増殖条件下での細胞中のタンパク質の局在」を参照)または組織サンプル(図「 組織サンプル中のタンパク質の局在」を参照)から、マーカータンパク質を容易かつ効果的に細胞コンパートメント特異的に局在化することができます。
See figures

Principle

キットには4つの抽出バッファーが含まれており、細胞溶解物や組織から細胞質基質、膜、核、細胞骨格に関連するタンパク質を連続的に分離できます。

Procedure

4つの抽出バッファーを細胞ペレットに連続的に添加し、それぞれの分画を遠心分離により分離します(フローチャートを参照)

組織用の別のプロトコールでは、細胞ペレットと同様に処理する前に、TissueRuptorを用いてサンプルのホモジェナイズと破砕を同時に行います。

Applications

タンパク質の細胞内分画は、存在量が少ない種の濃縮に使用できます。また、酵素、調節タンパク質、構造タンパク質の細胞内局在の定義、基礎条件下および刺激条件下での生体分子のコンパートメント再分布のモニタリングに使用できます。

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsSDS-PAGE、質量分析
Fractions isolated4分画
Start material細胞溶解物
Sample size~5 x 10e6細胞
Binding capacity/yield多様
Species真核生物

Resources

MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
キットハンドブック (2)
For the subcellular fractionation of proteomic
samples
クイックスタートプロトコール (1)
Safety Data Sheets (1)
Certificates of Analysis (1)
Kit Handbooks (2)
For the subcellular fractionation of proteomic
samples
Quick-Start Protocols (1)

FAQ

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
Do the Buffers CE1 - CE4 of the Qproteome Cell Compartment Kit contain DTT or any reducing agent?
No, none of the buffers of the Qproteome Cell Compartment Kit contain reducing agents.
FAQ ID -824
Will acetone precipitation recommended in the Qproteome Protocols denature Protein?
Acetone precipitation recommended in the protocols for the Qproteome Protein Fractionation Kits leaves most proteins in a native state. Some proteins may partially denature when precipitating with acetone.
FAQ ID -807
Is it possible to perform immunoprecipitation (IP) directly on protein fractions from the Qproteome Cell Compartment Kit?

We have not tested immunoprecipitation on protein fractions from the Qproteome Cell Compartment Kit. However, since Buffers CE1, CE2 and CE3 are native buffers, we would expect that they would work for this application. It might be a good idea to dilute the fractions 1:1 with a buffer already tested for immunoprecipitation to adjust to IP conditions. Buffer CE4 is strongly denaturing, so the final protein fraction 4 may not be used for immunoprecipitation.

 

FAQ ID -855
How do I perform an Acetone Precipitation for concentrating and desalting protein samples?

The following protocol is suitable for concentrating and desalting protein samples for downstream applications such as 2D-PAGE:

  • Add four volumes of ice-cold 100% acetone to the protein fraction and incubate for 15 minutes on ice
  • Centrifuge for 10 minutes at 12,000 x g in a pre-cooled microcentrifuge at 4°C
  • Discard the supernatant and air dry the pellet
  • Resuspend the pellet in an appropriate sample buffer required for the downstream application

You will find this protocol also in the handbooks for QIAGEN's QProteome Protein Fractionation Kits.

FAQ ID -1035
Which fraction will contain endosomal, microsomal and lysosomal proteins using the Qproteome Cell Compartment Kit?
Endosomal, microsomal and lysosomal proteins will be present in membrane protein fraction 2, as organelles remain intact after addition of Buffer CE1 during the first fractionation step with the Qproteome Cell Compartment Kit.
FAQ ID -808
Which fraction will contain soluble mitochondrial proteins using the Qproteome Cell Compartment Kit?
Soluble mitochondrial proteins will be present in membrane protein fraction 2 when using the Qproteome Cell Compartment Kit.
FAQ ID -839