Qproteome Cell Compartment Kit

用于根据细胞位置对蛋白进行分级分离

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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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Qproteome Cell Compartment 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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Features

  • 高重复性分离细胞蛋白组分
  • 可监测不同细胞生长状态下蛋白质的定位情况
  • 在进一步纯化和分析之前,根据亚细胞定位富集特定蛋白组分

Product Details

真核细胞是一个复杂、井然有序且高度结构化的系统。Cell Compartment Kit 设计用于快速、简便地对完好的真核细胞和组织进行亚细胞分级分离。在细胞团块中依次加入不同的提取缓冲液,可选择性地分离不同细胞区室中的蛋白。

Performance

使用 Qproteome Cell Compartment Kit 可以简单有效地对分离细胞中的标记蛋白进行细胞区室特异性定位(见图“ 标记蛋白的特异性分离”和“ 不同生长条件下细胞中蛋白质的定位”)或对组织样本中的标记蛋白进行同样的操作(见图“ 组织样本中蛋白的定位”)。
See figures

Principle

该试剂盒包含 4 种 Extraction Buffers,可从细胞裂解物或组织中依次分离与细胞溶质、细胞膜、细胞核和细胞骨架相关的蛋白。

Procedure

4 种 Extraction Buffers 依次加入细胞团块,然后通过离心分离出相应的馏分(见流程图)。

在组织的替代处理方案中,样本在像细胞团块一样接受处理之前,使用 TissueRuptor 同时进行均质化和破碎。

Applications

蛋白的亚细胞分离可用于富集低丰度物种;明确酶、调节蛋白和结构蛋白的亚细胞定位;以及监测生物分子在基础和刺激条件下的区室再分布。

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsSDS-PAGE、质谱法
Fractions isolated四个馏分
Start material细胞裂解物
Sample size约 5 x 10e6 个细胞
Binding capacity/yield不一
Species真核细胞

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (2)
For the subcellular fractionation of proteomic
samples
快速启动实验方案 (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