Active Aprotinin (AP)

BPTI; Trasylol; Pancreatic Trypsin Inhibitor; Basic protease inhibitor

ACTIVITY TEST

Aprotinin (AP) is a competitive serine protease inhibitor. Reversibly binds to and blocks the enzymatic active site. Inhibits a range of serine proteases including trypsin, chymotrypsin, kallikrein and plasmin. Inhibits cytopathogenic effect of SARS-CoV-2 and double-stranded RNA formation in SARS-CoV-2-infected cells. The activity of recombinant bovine AP was measured by its ability to inhibit trypsin cleavage of a peptide substrate BAPNA in the assay buffer 200 mM Triethanolamine hydrochloride, 20 mM CaCl2, pH 7.8. The reaction was performed in adding 20 μl 4 mg/mL trypsin diluted by 1mM HCl to 160 μl assay buffer and 20 ul 0.85% (w/v) NaCl and start the reaction by adding 100 µl of 1mg/ml BAPNA. Include a substrate blank containing 160 μl assay buffer, 20 μl 1mM HCl, 20 ul 0.85% (w/v) NaCl and 100 µL of 1mg/ml substrate. Rapidly mixing at 25 ℃, then read at 405 nm in kinetic mode for 5 minutes using a microplate reader controlling the ∆A405nm/min=0.08-0.12. The 20 ul different concentrations of recombinant bovine AP was incubated with 20 ul 4 mg/mL trypsin in 160 ul assay buffer at 25℃ for 10 minutes followed by adding 100 ul substrate, then read at 405 nm in kinetic mode for 5 minutes using a microplate reader. Under these conditions, the enzyme amount of 50% inhibition of trypsin activity per minute is defined as a unit. The specific activity of recombinant bovine AP is >3000 U/mg.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
24 Aprotinin prevents proteolytic epithelial sodium channel (ENaC) activation and volume retention in nephrotic syndrome. pubmed:29042083
Aus der Medizinischen Universitätsklinik diss_Menacher word
Acta Physiologica Plasminogen deficiency does not prevent sodium retention in a genetic mouse model of experimental nephrotic syndrome Pubmed: 32455507
Evaluation of The Relationship Between Apelin 36 and Oxidative Stress In Patients With General Anxiety Disorder
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