Preparation of lysozyme stock solution (10 mg/ml)

  • Lysozyme is an enzyme used for the extraction of proteins and nucleic acid from bacteria. It improves the extraction process by facilitating the lysis of bacterial cells by digesting the cell wall of bacteria.
  • It facilitates lysis of bacterial cells by degrading cell wall component peptidoglycans by catalyzing the hydrolysis of 1,4-beta-linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in peptidoglycans.
  • Lysozyme is a low molecular weight protein (14.4 kDa) which is composed of a single-chain polypeptide of 129 amino acids cross-linked with four disulfide bridges.
  • Lysozyme is commercially supplied as lyophilized powder which dissolves easily in aqueous buffer (e.g., Tris-HCl)
  • Since lysozyme loses its activity in solution, often a fresh solution is prepared.

Requirements:

Reagents and solutions
> Lysozyme (lyophilized powder)
> 10 mM Tris-HCl, pH 8.0

Equipment and disposables:
> Pipetman (P1000) and 1 ml tips
> 1.5 ml / 2ml Eppendorf tubes
> Analytical weighing balance

Composition
> 10 mg/ml lysozyme
> 10 mM Tris.Cl, pH 8.0

Objective
Preparation of 1 ml of lysozyme solution

Preparation

Step 1: Weigh out 10 mg lyophilized powder of lysozyme in an Eppendorf tube carefully to prepare a 1 ml solution.

Step 2: Add 1 ml of 10 mM Tris.Cl, pH 8.0 in it. Dissolve it by pipetting several times with a 1 ml pipette. 

Tip:
It is often difficult to weigh out exact amounts due to small quantities. In this case, weigh out an approximate quantity sufficient for your experiment and use solvent volume accordingly.

Storage
One can store the solution in the fridge for 1 month.

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