Beta Agarase Assay

The Worthington enzyme can be 2-4 times higher in activity than preparations offered by other suppliers. One Worthington unit will hydrolyze 500 milligrams of molten 1% LMP agarose per hour at 40-42°C.


  • 0.1 M Sodium acetate, pH 5.0, containing 0.001 M EDTA
  • 1% LMP Agarose, dissolved in 0.08 M Tris⋅borate buffer containing 0.001M EDTA, pH 8.3


To a series of tubes in 40-42°C water bath add 0.5 ml (500 mg gel) 1% LMP agarose gel.

Add 0.25 ml of buffer to each tube.

Prepare dilutions of the enzyme in 0.01% BSA/water to 1:5, 1:10, 1:20, 1:30, 1:40, and 1:50.

Add 10 ul of each dilution to a labelled tube. Add 10 ul of 0.01% BSA enzyme diluent to a tube and label as BLANK.

Incubate one hour at 40-42°C.

Remove from water bath and chill in ice bath for 5 minutes.

Inspect tubes for the last dilution having complete digestion. Complete digestion is indicated by liquefaction of the agarose. Use this tube to select the dilution factor and in the calculation of activity.



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