Qproteome Cell Compartment Kit

Para el fraccionamiento de proteínas según la ubicación celular

S_1084_5_GEN_disclaimer

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Qproteome Cell Compartment Kit

Cat. No. / ID:   37502

Para un máximo de 10 fraccionamientos subcelulares: Extraction Buffer, Protease Inhibitor Solution, Benzonase®.
6869,00 MXN
Log in To see your account pricing.
El Qproteome Cell Compartment Kit está concebido para su uso en aplicaciones de biología molecular. Este producto no está concebido para el diagnóstico, la prevención ni el tratamiento de enfermedades.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

Las células eucariotas son sistemas complejos, bien ordenados y muy estructurados. El Cell Compartment Kit está diseñado para el fraccionamiento subcelular rápido y sencillo de células y tejidos eucariotas intactos. Por una adición secuencial de distintos tampones de extracción al sedimento de la célula, las proteínas de los distintos compartimentos celulares se pueden aislar selectivamente.

Performance

El uso del Qproteome Cell Compartment Kit permite una localización sencilla y eficaz de proteínas marcadoras específicas de compartimentos celulares a partir de células aisladas (consulta la figura « Separación específica de proteínas marcadoras» y « Localización de proteínas en células en diferentes condiciones de crecimiento») o de muestras de tejido (consulte la figura « Localización de proteínas en muestras de tejido»).
See figures

Principle

El kit contiene 4 Extraction Buffers que permiten el aislamiento secuencial de las proteínas asociadas con el citosol, las membranas, el núcleo y el citoesqueleto a partir de lisados o tejidos celulares.

Procedure

Los 4 Extraction Buffers se añaden secuencialmente a un sedimento celular y las fracciones respectivas se aíslan por la centrifugación (consulte el diagrama de flujo).

En un protocolo alternativo para los tejidos, las muestras se homogenizan al mismo tiempo y se separan con el TissueRuptor antes de procesarse por sedimento celular.

Applications

El fraccionamiento celular de proteínas se puede utilizar para el enriquecimiento de especies de baja abundancia, para definir la localización subcelular de las enzimas, proteínas reguladoras y estructurales y para supervisar la redistribución compartimental de biomoléculas en condiciones basales y estimuladas.

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsSDS-PAGE, espectrometría de masas
Fractions isolatedCuatro fracciones
Start materialLisado celular
Sample size~ 5 x 10e6 células
Binding capacity/yieldVaría
SpeciesEucariotas

Resources

Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
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