Lysis Buffers for Protein Extraction

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

Search RIPA buffers online.

 

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

Search Minute™ extraction kits online.

 

Protease inhibitors

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.

 

Serine proteases
AEBSF	Hydrochloride >98%	401070	100mg
		401071	1g
Aprotinin	Powder, bovine lung 6300 KIU/mg, 3.5 pH.Eur.U/mg	BXH470	100mg
	Solution, 200 000KIU/mL – Pharma Grade	24431A	50mL
PMSF	>99,9%	147376	5g
		24431A	50mL
Benzamidine	Hydrochloride, anhydrous, >99%	003055	100g
Metalloproteases
EDTA	Disodium Salt >99.5%, Molecular Biology Grade	036290	1kg
EGTA	>99 %	10075A	25g
1,10-Phenanthroline	Anhydrous, >99 %	E4414E	25g
Cysteine proteases
E-64	>99%	789581	5mg
		789582	25mg
Iodoacetamide	>99 %	020483	25g
Leupeptin	Hemisulfate >97%, Synthetic	201736	25mg
NEM	N-Ethylmaleimide >98 %	04474B	25g
Aspartate proteases
Pepstatin	>90 %	827752	25mg
Protease inhibitor cocktails
Protease Inhibitor Cocktail I General use (AEBSF Hcl 500µM, Aprotinin 150nM, E-64 1µM, EDTA 0,5mM, Leupeptin 1µM)		WT0900	1 flacon
Protease Inhibitor Cocktail II for Bacteria (AEBSF Hcl 20mM, Bestatin 1,7mM, E-64 200µM, EDTA 85mM, Pepstatin A 2mM)		WT8260	1 flacon

 

Detergents

Detergents are used to separate membrane proteins from membrane and to open intracellular compartments like lysosomes. They can stabilise some proteins in hydrophilic environment.

 

Nondenaturing
CHAPS	>98 %	33351A	10g
Triton X-100	10% solution, Carbonyl and peroxide free (>99%)	BT84L0	50mL
	10 % solution, Oxidant-free	521121	5x10mL
Denaturing
SDS	20 % solution, 0.2 micron filtered, MB grade	896826	500mL

 

Others

Molecular Biology Grade (DNase, RNase free)
Guanidine HCl	>99.5 %, chaotropic agent	018380	500g
D-Glucose	Anhydrous, >99.9%	40250B	500g
Glycerol	>99.8%, Sterile	047624	500mL
EDTA	disodium salt, >99.5%	036290	1kg
DTT	>99 %	054721	5g
Urea	>99.5%, Ammonia-, Cyanate-free	031904	1kg
Tris	1M pH 7.5, Sterile solution	N13713	1L