What is pepstatin used for?
Pepstatin, a reversible peptidomimetic inhibitor of aspartic peptidases, inhibits cathepsin D with Ki in pM range [34]. It is a valuable research tool for in vitro and in vivo regulation of mature cathepsin D.
How do you dissolve Leupeptin?
Leupeptin: Dissolve 50 mg in 10.5 ml of water to get 10 mM stock (1000X). Working concentration is 10 µM, so add 1µl of stock in 1 ml of lysis buffer. Pepstatin: Dissolve 5 mg in 7.3 ml of Methanol or DMSO to get 1 mM stock (1000X).
What does pepstatin do to pepsin?
Pepstatin, a specific inhibitor of acid proteases, binds tightly to pepsin. Although the binding is not of the covalent nature, the inhibition roughly follows the stoicheometrical mode. Pepstatin can be used to titrate pepsin. Formation of an equimolar pepsin-pepstatin complex can be shown by gel filtration.
How do you dissolve pepstatin A?
It has been dissolved at 10 mg/mL in ethanol with heat. The resulting solution is colorless, but may appear hazy. To remove haziness, add up to 50 µl of glacial acetic acid per mL of ethanol. At 25 mg/mL DMSO Pepstatin A forms a clear, faint yellow solution.
How do you dissolve pepstatin?
Pepstatin A is only sparingly soluble in water. 4 It is normally dissolved in a solvent such as ethanol, methanol, or DMSO and diluted into buffer. The stock solution should be of a concentration that the solvent used is diluted at least 1000X in the working solution.
How do you dissolve protease?
Dissolve one tablet in 50 ml aqueous buffer or water. If very high proteolytic activity is present, use one tablet for 25 ml buffer. Dissolve one tablet in 10 ml aqueous buffer or water. If very high proteolytic activity is present, use one tablet for 7 ml buffer.
How is pepsin inhibited?
Inhibitors. Pepsin may be inhibited by high pH (see Activity and stability) or by inhibitor compounds. Pepstatin is a low molecular weight compound and potently inhibitor specific for acid proteases with a Ki of about 10−10 M for pepsin.
What type of inhibitor is Benzamidine?
competitive inhibitor
Benzamidine is a reversible competitive inhibitor of trypsin, trypsin-like enzymes and serine proteases.
How do you make a protease inhibitor solution?
Protease Inhibition 3. Add 10 μl of 100mM Benzamidine HCl (see Benzamidine HCl stock solution protocol) per 1 ml of lysis buffer. 4. Add 10 μl of 10mM Leupeptin Hemisulfate (see Leupeptin stock solution protocol).
How do you inhibit protease activity?
These molecules inhibit protease activity through an allosteric mechanism. BBI, a trypsin inhibitor from soybeans [14] and aminoglycosides, inhibitors of the anthrax lethal factor protease [15] are examples of non-competitive inhibitors.
How do you get rid of pepsin in the airways?
It is recommended to take a proton pump inhibitor in the morning, and avoid eating or drinking for 20 minutes. Also avoiding carbonated beverages, tomato-based products, citrus products, spicy foods, chocolate, breath mints, coffee, caffeinated beverages and alcohol reduces the activation of pepsin.
Is Benzamidine a competitive inhibitor?
Benzamidine is a reversible competitive inhibitor of trypsin, trypsin-like enzymes and serine proteases.