Native lysis uses mild non-ionic detergents such as NP-40 or Triton X-100. It maintains protein folding and interactions, which is needed for co-immunoprecipitation and enzyme activity assays. Denaturing lysis uses SDS or urea to fully solubilise all proteins, regardless of hydrophobicity. This is preferred for maximal protein recovery in Western blot and mass spectrometry workflows, where native conformation is not required.
Sample Preparation
Protein Purification Kits & Reagents
Protein extraction and subcellular fractionation kits for clean, unbiased samples
Spin-column-based total protein extraction for protein purification uses a proprietary resin to recover proteins across a broad molecular weight range. This gives you clean starting material for proteomics, structural studies, or functional assays, regardless of extraction method. The column format lyses cells or tissues in under one minute, without sonication or RIPA buffer. To break up tissue before lysis, pair these kits with the Precellys tissue homogenisers.
This approach eliminates non-systematic protein loss and helps preserve post-translational modifications when appropriate protease and phosphatase inhibitors are used. It delivers a complete, unbiased protein profile compatible with Western blot, mass spectrometry, and protein quantitation assays.
Subcellular fractionation kits isolate specific compartments, including plasma membrane, cytosolic, mitochondrial, and nuclear fractions, without ultracentrifugation. Formats are available for animal cells, solid tissues, plant material, and specialised matrices such as blood vessels. Organelle isolation kits for yeast nuclei, yeast mitochondria, plasma membranes, and lysosomes are validated for small starting material, accounting for the rigid cell wall.
Nuclear extraction protocols use non-denaturing conditions to preserve protein-protein interactions. Protease inhibitor cocktails protect sample integrity during lysis, and protein stabilisation reagents extend storage stability without freeze-thaw degradation. These protein purification kits cover workflows from total cell lysate through organelle-level fractionation.
Applications
Western blot and co-immunoprecipitation
Native lysis preserves protein-protein interactions and post-translational modifications, so it suits co-IP and proximity ligation. Pair it with validated primary antibodies for detection.
Mass spectrometry proteomics
Denaturing lysis with SDS or urea maximises solubilisation. Filter-aided sample preparation then removes the detergent before tryptic digestion.
Subcellular and organelle fractionation
Isolate plasma membrane, cytosolic, mitochondrial, and nuclear fractions without ultracentrifugation. Formats cover animal cells, solid tissue, plant material, and yeast.
Enzyme activity and functional assays
Fractionation recovers mitochondrial or nuclear proteins in a functional state. This lets you measure organelle-specific enzyme activity in the right compartment.
Structural biology and biophysics
X-ray crystallography, cryo-EM, and SPR need high-purity, aggregate-free preparations. Start from purified recombinant proteins for these workflows.
Protein quantification and normalisation
Measure total protein before gel loading, ELISA normalisation, or proteomics. Match your extract to a protein quantification assay compatible with your buffer.
Frequently asked questions
Cell lysis releases endogenous proteases stored in lysosomes and cytoplasm. These begin degrading proteins within minutes at room temperature. Broad-spectrum inhibitor cocktails typically include serine protease inhibitors (PMSF or AEBSF), cysteine protease inhibitors (E-64), and metalloprotease inhibitors (EDTA or bestatin). Omitting inhibitors leads to truncated bands and reduced yield on Western blots, particularly for low-abundance or labile targets.
Yeast have a rigid cell wall composed of glucan and chitin that resists detergent-based lysis. Effective disruption requires mechanical methods, such as glass bead beating or freeze-thaw cycles with liquid nitrogen. You can also use enzymatic removal of the cell wall with zymolyase or lyticase before detergent lysis. Mammalian cells lack this wall and lyse readily with mild detergent buffers, making bead beating unnecessary for routine mammalian protein extraction.
Most spin-column total protein extraction formats are validated for 1 to 30 mg of tissue, or 10,000 to 5 million cells per column. For rare or precious samples, starting with a minimum of 5,000 cells can still yield sufficient protein for a Western blot, when the target is moderately abundant. Scaling buffer volumes proportionally and avoiding over-loading the column membrane are the two most critical adjustments at low input.
Product catalogs
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