The proteomic analysis starts with lysis, followed by purification. Lysis buffers include detergents and other reagents for achieving efficient protein solubility. These article presents a selection of commonly used lysis buffer, kit and related biochemicals.
The diversity of source samples and proteins does not allow us to offer an universal solution. A first possibility is to try the ready-to-use buffers known to work on the majority of organisms. Then there are specialized kits for certain proteins or like the Minute™ kit above, optimized for organisms such as bacteria.
To improve lysis according to specific needs, we offer a complete set of protease inhibitors, detergents and other high purity biochemicals, dedicated to biological applications.
RIPA lysis and Extraction Buffer
For the Optimal Lysis of Adherent & Suspension Cells – compatible with purification
|A complete lysis buffer for the release of cytoplasmic, membrane and nuclear proteins from adherent and suspension cultured mammalian cells. The RIPA lysis buffer is fully compatible with many applications, including reporter assays, protein assays (BC Assay), immunoassays and other protein purification techniques. It is preferred for nuclear, cytoplasmic and mitochondrial proteins, whereas cytoskeletal and extracellular region proteins are more soluble in urea.|
RIPA Lysis Buffer does not contain protease inhibitors, however it is fully compatible with a range of individual protease inhibitors and cocktails.
|Protein digestion||Mass Spectrometry based protein identification
and analysis (HPLC-MS)
RIPA lysis and Extraction Buffer HG4361 100mL
Contents 25mM Tris, 150mM Sodium Chloride, 1% NP-40, 1% Sodium Deoxycholate, 0.1% SDS, pH 7.6
Minute™ Bacterial Total Protein Extraction Kit
To rapidly extract total proteins from cultured bacterial cells for applications such as SDS-PAGE, immunoblotting, ELISA and other applications.
|Minute™ Bacterial Total Protein Extraction Kit is composed of optimized denaturing cell lysis buffers and protein extraction filter cartridges with 2.0 ml collection tubes. The kit is designed to quickly extract denatured proteins from bacteria. It contains sufficient materials for the extraction of total proteins from a 100 ml E. coli culture.|
Comparison of protein profiles extracted by Minute kit and RIPA buffer.
Lanes 1, 2 and 3 are proteins extracted by Minute kit, RIPA buffer and insoluble fraction of RIPA extraction respectively.
Minute™ Bacterial Total Protein Extraction Kit SB-004 50 preps
Contents 25mL bacterial lysis buffer A + 2.5mL buffer B + 50 protein extraction filter cartridges (500µL)+ 50 collection tubes with cap
|Protease inhibitors are molecules that inhibit protease function by binding either reversibly or irreversibly to the enzyme. Most inhibitors are only active against one or two types of proteases.|
|Aprotinin||Powder, bovine lung 6300 KIU/mg, 3.5 pH.Eur.U/mg||BXH470||100mg|
|Solution, 200 000KIU/mL – Pharma Grade||24431A||50mL|
|Benzamidine||Hydrochloride, anhydrous, >99%||003055||100g|
|EDTA||Disodium Salt >99.5%, Molecular Biology Grade||036290||1kg|
|1,10-Phenanthroline||Anhydrous, >99 %||E4414E||25g|
|Leupeptin||Hemisulfate >97%, Synthetic||201736||25mg|
|NEM||N-Ethylmaleimide >98 %||04474B||25g|
Protease inhibitor cocktails
|Protease Inhibitor Cocktail I
General use (AEBSF Hcl 500µM, Aprotinin 150nM, E-64 1µM, EDTA 0,5mM, Leupeptin 1µM)
|Protease Inhibitor Cocktail II
for Bacteria (AEBSF Hcl 20mM, Bestatin 1,7mM, E-64 200µM, EDTA 85mM, Pepstatin A 2mM)
|Detergents are used to separate membrane proteins from membrane and to open intracellular compartments like lysosomes. They can stabilise some proteins in hydrophilic environment.|
|Triton X-100||10% solution, Carbonyl and peroxide free (>99%)||BT84L0||50mL|
|10 % solution, Oxidant-free||521121||5x10mL|
|SDS||20 % solution, 0.2 micron filtered, MB grade||896826||500mL|
Molecular Biology Grade (DNase, RNase free)
|Guanidine HCl||>99.5 %, chaotropic agent||018380||500g|
|EDTA||disodium salt, >99.5%||036290||1kg|
|Urea||>99.5%, Ammonia-, Cyanate-free||031904||1kg|
|Tris||1M pH 7.5, Sterile solution||N13713||1L|