Proteases and Peptidases

Substrates. Protein substrates for proteases are most often hemoglobin or casein and must be completely soluble in buffer. Casein for protease assays is designated "nach Hammarsten," while hemoglobin "for protease assay" is usually of high quality. Casein precipitates below pH 6, so it is used at neutral to alkaline pH. Hemoglobin must be denatured before use, either by treatment with acid (if the assay is at acidic pH) (16) or urea (neutral to alkaline pH assay) (17). Gelatin, sometimes used in viscometric assays, is quite heterogeneous, so lot-to-lot reproducibility is a con-

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