Qproteome Mitochondria Isolation Kit

고순도 미토콘드리아 제제의 효율적인 분리에 사용

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Qproteome Mitochondria Isolation Kit

Cat. No. / ID:   37612

12가지 고순도 미토콘드리아 제제용 완충액 및 시약
Qproteome Mitochondria Isolation Kit은/는 분자생물학 분야에 사용하기 위한 것입니다. 이 제품은 질병의 진단, 예방, 또는 치료용이 아닙니다.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 고순도, 무손상 미토콘드리아
  • 표준화된 절차를 사용하여 향상된 재현성
  • 편리하고 빠르며 사용하기 쉬운 키트 형태

Product Details

Qproteome Mitochondria Isolation Kit는 표준 작업대용 원심분리기를 사용하는 빠르고 재현 가능하며 표준화된 절차에서 매우 높은 순도의 완전 활성 미토콘드리아 제제를 제공합니다.

Performance

문헌의 차등 원심분리 방법과 대조적으로, 세포 구획 표지자 단백질 분석에 따르면 Qproteome Mitochondria Isolation Kit는 교차 오염이 거의 없는 미토콘드리아 분획을 제공합니다(그림 '교차 오염이 거의 없는 미토콘드리아 분획' 참조). 세포질 오염 물질이 거의 없기 때문에 2D-PAGE 분석에서와 같이 낮은 함량의 종을 쉽게 분석할 수 있고, 원활한 절차를 통해 무손상 미토콘드리아가 포함된 고농축 분획을 제공합니다(그림 ' 미토콘드리아 막이 손상되지 않은 고순도 미토콘드리아 ' 참조). 무결성은 형광 염료의 흡수량을 측정하여 입증할 수도 있습니다(그림 '단백질 1mg당 추가 무손상 미토콘드리아' 참조).
See figures

Principle

미토콘드리아를 연구하는 연구자들이 직면하는 주요 문제 중 하나는 미토콘드리아와 세포질 분획을 효율적으로 분리하는 것입니다. 기존의 준비 방법은 지루하고 시간이 많이 걸리며 초원심분리기를 이용해야 합니다. Qproteome Mitochondria Isolation Kit는 표준 작업대용 원심분리기를 사용하는 빠르고 재현 가능하며 표준화된 절차에서 매우 높은 순도의 완전 활성 미토콘드리아 제제를 제공합니다.

Applications

미토콘드리아는 세포자멸, 단백질체학, 생물 의학 연구, 신약 개발 등 수많은 분야에서 집중적으로 연구하는 주제입니다. Qproteome Mitochondria Isolation Kit는 단백질이 고유의 형태와 완전한 생물학적 활성을 유지하는 고순도 미토콘드리아 단백질을 제공하므로 세포자멸 연구, 2D-PAGE 분석, 기타 생화학적 분석에 적합합니다.

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsKOSDS-PAGE, 질량 분석법, 활동 분석
Sample size5 x 10e6 - 2 x 10e7 cells, 20 mg - 60 mg
For glycoproteins: which type of glycoproteinsn.d
Binding capacity/yield20~80µg
Species진핵
Fractions isolated분획 1개
Start material세포, 조직

Resources

Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Kit Handbooks (1)
For purification of mitochondria from eukaryotic cells and tissues
Certificates of Analysis (1)

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