Qproteome Cell Compartment Kit

세포 위치에 따른 단백질 분획에 사용

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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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₩549,000.00
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Qproteome Cell Compartment Kit은/는 분자생물학 분야에 사용하기 위한 것입니다. 이 제품은 질병의 진단, 예방, 또는 치료용이 아닙니다.

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Features

  • Cytosol, membrane, nucleus, cytoskeleton에 존재하는 protein을 효율적으로 분리
  • 다른 cell culture 조건에서 단백질 localization monitoring
  • Cell compartment-specific proteins for further purification or analysis

Product Details

진핵 세포는 복잡하고 잘 정렬되어 있으며 고도로 구조화된 시스템입니다. Cell Compartment Kit는 손상되지 않은 진핵 세포 및 조직의 쉽고 빠른 세포하 분획을 위해 설계되었습니다. 세포 펠릿에 다양한 추출 완충액을 순차적으로 추가하면 다양한 세포 구획에서 단백질을 선택적으로 분리할 수 있습니다.

Performance

Qproteome Cell Compartment Kit를 사용하면 분리된 세포(그림 ' 표지자 단백질의 특이적 분리' 및 ' 다양한 성장 조건에서 세포 내 단백질의 국소화' 참조) 또는 조직 샘플(그림  조직 샘플 내 단백질의 국소화' 참조)에서 표지자 단백질의 세포 구획별 국소화를 간단하고 효과적으로 수행할 수 있습니다.
See figures

Principle

이 키트에는 세포 용해물 또는 조직에서 세포질, 막, 핵, 세포골격과 관련된 단백질을 순차적으로 분리할 수 있는 추출 완충액 4개가 포함되어 있습니다.

Procedure

추출 완충액 4개를 세포 펠릿에 순차적으로 추가하고 각 분획을 원심분리로 분리합니다(순서도 참조).

조직에 대한 대체 프로토콜에서 샘플은 세포 펠릿으로 처리되기 전에 TissueRuptor를 사용하여 동시에 균질화되고 파괴됩니다.

Applications

단백질의 세포하 분획은 낮은 함량의 종을 농축하고, 효소, 조절 단백질, 구조 단백질의 세포하 국소화를 정의하고, 기초 및 자극 조건에서 생체분자의 구획 재분포를 모니터링하는 데 사용할 수 있습니다.

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsSDS-PAGE, 질량 분석법
Fractions isolated분획 4개
Start material세포 용해물
Sample size~5 x 10e6 cells
Binding capacity/yield다양함
Species진핵

Resources

Safety Data Sheets (1)
Kit Handbooks (2)
For the subcellular fractionation of proteomic
samples
Quick-Start Protocols (1)
Certificates of Analysis (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