Protease and protease inhibitors are all-enerdy.

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< The release of proteases, > which need to be quickly inhibited to keep the protein from degradation, is released at the same time as < a href> is extracted from broken cells

During protein extraction, it is necessary < to add > protease inhibitors to prevent protein hydrolyzing. The following are five commonly used protease inhibitors and their respective characteristics, as the sensitivity of various proteases to different proteins varies, so the concentration of various proteases needs to be adjusted. Since protease inhibitors are extremely soluble in liquids, particular attention should be paid to the addition of human protease inhibitors to the buffer to reduce the precipitation of protease inhibitors. A more complete list of proteases and protease inhibitors can be found in Paulingman's catalog.

commonly used inhibitors

PMSF

1) inhibit serine proteases (e.g. pancreatic lactase, trypsin, clotting enzymes) and -based proteases (e.g.

2) 10mg/ml dissolved in isopropyl

3) can be kept at room temperature for

4) Working concentration: 17 to 174ug/ml (0.1

5) Unstable in a water liquid solution, fresh PMSF must be added to each separation and purification step.

< a href" > EDTA

1) inhibits metal protein hydroly

2) 0.5mol/L aqueous solution, pH8 to 9;

3) The solution is stable at 4 degrees C for more than

4) Working concentration: 0.5 to 1.5mmol/L. (0.2

5) Add NaOH to adjust the pH of the solution, otherwise EDTA insoluble.

protease inhibitors (pepstantin)

l) inhibit acid proteases such as gastroprotease, angiotensin pep >tidease,protease D and coagulase;

2) 1mg/ml dissolved

3' storage fluid is stable within 4 oC for 6

4)1 concentration: 0.7ug/ml (1umol/L

5) insoluble in water.

leupeptin

1) inhibits serine and -based proteases, such as papaya protease, plasma enzyme and tissue protease B;

2) lOmg/ml is soluble in

3) The storage fluid is stable for one week at 4 degrees C and at -20 degrees C

4) working concentration of 0.5 mg/ml.

aprotinin

1) inhibits serine proteases, such as plasmaase, vasosorrene, trypsin and pancreatic

2) lOmg/ml soluble in water, pH7 to 8

3) storage fluid 4 degrees C stable for one week, -20 degrees C stable for 6

4) working concentration: 0.06 to 2.0ug/ml (0.01

5) Avoid repeated freezing and melting:

6) institation at pH.gt;12.8.

35ug/ ml PMSF for a

of protease inhibitors... Serine protease inhibitors

0.3mg/ ml EDTA ... Metal protease inhibitors

0.7ug/ml gastric protease inhibitors (Pepstatin)... Acid protease inhibitors

0.5ug/ml bright proteoproteinase (Leupeptin)... Broad-spectrum protease inhibitors

commonly used protease inhibitors table