What is pepstatin?

Anonymous

∙ 16y ago

Updated: 8/16/2019

Pepstatin A is an inhibitor of acid proteases (aspartyl peptidases). It forms a 1:1 complex with proteases such as pepsin, renin, cathepsin D, bovine chymosin, and protease B (Aspergillus niger). The inhibitor is highly selective and does not inhibit thiol proteases, neutral proteases or serine proteases. Solublized Beta-secretase and retroviral protease are also inhibited by Pepstatin A. It has been used to characterize proteases from several sources. Pepstatin A is thought to inhibit by a collected-substrate inhibition mechanism.

Wiki User

∙ 16y ago

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How much of each ingredient do I need to make a 1mL lysis buffer solution?

You need to specify, lysis buffer for bacteria or eukaryotic cell. The most common buffer cocktail for eukaryotic cells is composed by an hypertonic solution and cell disrupters. To answer this question the proper amounts of the ingredients for 1 mL would be: 8.77 mg of NaCl (150 mM, the hypertonic component) 10 microliters NP-40 [can be replaced by 1 microliter (0.1%) of Triton X-100, a detergent] 6.06 mg of Tris in 100 microliters [Tris hydroxymethyl aminomethane, make 10 mL of a 100X solution (that is, 605.5 mg in 10 mL water, adjust to pH 8.0)] 890 microliters of distilled water. In some cases can be added some protease inhibitors such as PMSF, leupeptin, Aprotinin, etc. at concentrations of 1 microgram/mL. Store at 4oC for one month. Now, to make a lysis buffer for bacteria, the composition is different. For 1 mL mix: 6.06 mg of Tris in 100 microliters, as before 17.53 mg of NaCl (300 mM) 100 microliters of PMSF [Phenylmethylsulfonyl Fluoride, make 10X solution (1.742 mg in 10 mL of water)] 980 microliters of distilled water add aprotinin, leupeptin, and pepstatin at final concentration of 1 microgram/mL. Usually, to break the bacteria cell wall it is useful a lab blender. Disruption of both, eukaryotic and bacteria cells, must be done in cold conditions, usually on ice-water bath.

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What is pepstatin?

What else can I help you with?

How much of each ingredient do I need to make a 1mL lysis buffer solution?

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