Qproteome Nuclear Protein Kit

For separation of nuclear and nucleic acid binding proteins

Features

  • Nuclear preparations free of cytosolic proteins
  • Greatly reduced sample complexity
  • Highly reproducible fractionation
  • Includes protocols for tissue proteomics
Qproteome Nuclear Protein Kit

Cat. No. / ID: 37582

For 12 nuclear protein preparations: Buffers, Reagents, Protease Inhibitor Solution, Benzonase
The Qproteome Nuclear Protein Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

The Qproteome Nuclear Protein Kit is designed for specific enrichment of nuclear proteins from cultured mammalian cells.

Performance

The Qproteome Nuclear Protein Kit provides highly effective and reproducible separation of nuclear proteins as demonstrated using specific marker proteins (see figure " Reproducible, efficient separation of marker proteins"), which remain functionally active (see figure " Isolation of an active transcription factor").
See figures

Principle

The Qproteome Nuclear Protein Kit is designed for specific enrichment of nuclear proteins from cultured mammalian cells. Lysis and centrifugation are used to separate the cytosolic fraction (supernatant) from the cell nuclei (pellet). A high-salt buffer allows dissociation of nuclear binding proteins (such as transcription factors) and their removal by diffusion from the nuclei.

Procedure

Cells are lysed and centrifuged to isolate nuclei. After washing, an extraction buffer is added to the nuclei and nucleic acid binding proteins dissociate from DNA and RNA. This soluble fraction is separated from the nuclear pellet by centrifugation. Proteins remaining in the pellet are solubilized using a second extraction buffer (see figure " Nuclear protein fractionation procedure").
See figures

Applications

The Qproteome Nuclear Protein Kit delivers a nucleic acid binding protein fraction suitable for a wide range of activity assays.

Supporting data and figures

Specifications

FeaturesSpecifications
applicationsSDS-PAGE, mass spectrometry
fractionsisolatedThree fractions
speciesMammal
samplesize5 x 10e6 cells
forglycoproteinswhichtypeofglycoproteinsn.d
bindingcapacityyield200–300 µg
startmaterialCell cultures

Resources

Kit Handbooks (1)
For isolation of nuclear and nucleic acid binding proteins from eukaryotic cells and tissues

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
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