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Qproteome Bacterial Protein Prep Kit

For preparation of soluble bacterial proteins

Features

  • Gentle but effective cell lysis, high yields of active proteins
  • Simple procedure, no specialized equipment required
  • Highly reproducible procedure, consistent results time after time

Product Details

The kit contains a lysis buffer for preparation of soluble proteins from bacterial cultures for use in proteomics procedures.

Principle

Bacterial cell walls are disrupted by the combination of a freeze–thaw cycle and a lysis buffer containing lysozyme. The lysis buffer also contains a nuclease, which reduces the viscosity of the lysate and frees proteins associated with nucleic acids. Once lysis is complete, the lysate is centrifuged to precipitate insoluble proteins and cell debris.

Procedure

Lysis buffer is added to frozen cell pellets, cells are resuspended, and incubated on ice for 30 minutes. The lysate is centrifuged to precipitate insoluble proteins and cell debris, and soluble proteins are collected in the supernatant.

Applications

The Qproteome Bacterial Protein Prep Kit is used for preparation of soluble bacterial proteins for proteomics procedures (e.g., 2D-PAGE, mass spectrometry).

Specifications

FeaturesSpecifications
applicationsSDS-PAGE, mass spectrometry
samplesizePellet from a 250 ml sample
startmaterialCell cultures
speciesBacteria
fractionsisolatedOne fraction

Resources

Kit Handbooks (1)
For preparation of soluble proteins from
bacterial cell cultures

FAQ

Is it possible to adjust the Lysis Buffer volume used with the Qproteome Bacterial Protein Preparation Kit?

Yes, scaling the volume of the Native Lysis Buffer of the Qproteome Bacterial Protein Prep Kit is possible. Please adjust the bacterial culture volume proportionally, i.e. use 40 ml of Lysis Buffer for 1 liter of bacterial culture, or use 1 ml of Lysis Buffer for 25 ml of bacterial culture.

FAQ ID -841
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