Table 1.

Commonly used reagents in lysis buffer and their concentration

Common lysis buffers	Common reagents in lysis buffer	Typical concentration range	Role	References
NP-40 buffer, RIPA buffer, Tris-Triton buffer, Tris-HCl buffer	Tris-HCl	50 mM-100 mM (pH 7.4 –8.0)	pH buffering of solution	[5,8,22]
	NaCl	50 mM− 150 mM	Cell membrane rupture and protein solubility	[5,8,22]
	Non-ionic detergents: NP40 Octyl Glucoside Triton X-100 Tween	0.1–2%	Solubilization of non-polar insoluble proteins	[5,8,19,22,97]
	Ionic Detergents: SDS Sodium deoxycholate CTAB	0.1–1% 0.1–0.5% 0.01–0.5%	Coat proteins with negative charge and facilitates their separation on gels based on their molecular mass Membrane disruption Solubilization of poorly soluble proteins	[5,8]
	Zwitterionic CHAPS (SB3–10 and ASB-14)	2%	Break protein-protein interactions	[98,99]
	Urea	5–8M	Breaks non-covalent interactions and leads to protein denaturation	[5,99]
	EDTA	1–5 mM	Chelate metal ions, reduce oxidation damage and inhibition of metalloproteases	[8]
	EGTA		Chelate metal ions	[5]
	Glycerol	5–10%	Stabilization	[5,97]
	DTT	1–10 mM	Cleavage of disulfide bonds and protein denaturation	[5,97,99]
	2-Mercaptoethanol ^	1–10 mM	Cleaves disulfide bonds and protein denaturation	[5,97]
	TCEP	10–50 mM	Cleaves disulfide bonds destabilizes the secondary and tertiary structure	[97]
	Na3VO4	0.1– 2 mM	Tyrosine and alkaline phosphatase inhibition	[5,19]
	NaF	50 mM	Serine/threonine phosphatase inhibition	[5]
	Protease inhibitor cocktail	Usually in prepackaged tablet or liquid forms	Inhibits proteolysis	[5,8]

Abbreviations: Sodium dodecyl sulphate (SDS); Nonidet- 40 (NP-40); Radio immunoprecipitation assay (RIPA); Ethylene diamine tetra acetic acid (EDTA); Ethylene glycol tetra acetic acid (EGTA); Dithiothreitol (DTT); Sodium vanadate (Na₃VO4); Sodium fluoride (NaF); cetyl trimethyl-ammonium bromide (CTAB) (CTAB); (tris(2-carboxyethyl)phosphine (TCEP).