Bioactivity
(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 pig 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 ul 4 mg/mL trypsin diluted by 1mM HCl to 160 ul assay buffer and 20 ul 0.85% (w/v) NaCl and start the reaction by adding 100 ul of 1mg/ml BAPNA. Include a substrate blank containing 160 ul assay buffer, 20 ul 1mM HCl, 20 ul 0.85% (w/v) NaCl and 100 uL of 1mg/ml substrate. Rapidly mixing at 25 degree C, 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 pig AP was incubated with 20 ul 4 mg/mL trypsin in 160 ul assay buffer at 25 degree C 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 pig AP is >600 U/mg.)
Aprotinin (AP) Active Protein | AP active protein
Active Aprotinin (AP)
Applications
Activity Assay, Cell Culture
Purity
Greater than 90% by SDS-PAGE
Synonyms
Aprotinin (AP); N/A; Active Aprotinin (AP); Aprotinin; AP active protein (eukaryotic,293F cell), Pig; BPTI; Trasylol; Pancreatic Trypsin Inhibitor; Basic protease inhibitor4; AP active protein
Host
293F cell
Purity/Purification
Greater than 90% by SDS-PAGE
Form/Format
Freeze-dried powder; PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300.
Sequence
Cys24~Asp128
Applicable Applications for AP active protein
Activity Assay, Cell Culture
Species
Pig
Expression System
Eukaryotic expression
Tag
N-terminal His Tag and C-terminal Fc Region of Human IgG1
Endotoxin Level
<1.0EU per 1ug (determined by the LAL method)
Isoelectric Point
4.7
Research Area
Enzyme & Kinase, Cardiovascular biology, Human gene deletion, Hepatology
Preparation and Storage
Store at 2-8 degree C for one month, -80 degree C for 12 months
Bioactivity
(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 pig 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 ul 4 mg/mL trypsin diluted by 1mM HCl to 160 ul assay buffer and 20 ul 0.85% (w/v) NaCl and start the reaction by adding 100 ul of 1mg/ml BAPNA. Include a substrate blank containing 160 ul assay buffer, 20 ul 1mM HCl, 20 ul 0.85% (w/v) NaCl and 100 uL of 1mg/ml substrate. Rapidly mixing at 25 degree C, 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 pig AP was incubated with 20 ul 4 mg/mL trypsin in 160 ul assay buffer at 25 degree C 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 pig AP is >600 U/mg.)
Bioactivity
(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 pig 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 ul 4 mg/mL trypsin diluted by 1mM HCl to 160 ul assay buffer and 20 ul 0.85% (w/v) NaCl and start the reaction by adding 100 ul of 1mg/ml BAPNA. Include a substrate blank containing 160 ul assay buffer, 20 ul 1mM HCl, 20 ul 0.85% (w/v) NaCl and 100 uL of 1mg/ml substrate. Rapidly mixing at 25 degree C, 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 pig AP was incubated with 20 ul 4 mg/mL trypsin in 160 ul assay buffer at 25 degree C 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 pig AP is >600 U/mg.)
SDS-PAGE
(Figure. SDS-PAGE)
SDS-PAGE
(Figure. SDS-PAGE)
Product Categories/Family for AP active protein
NCBI and Uniprot Product Information
Similar Products
Product Notes
The AP (Catalog #AAA161813) is an Active Protein produced from 293F cell and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Aprotinin (AP) can be used in a range of immunoassay formats including, but not limited to, Activity Assay, Cell Culture. Researchers should empirically determine the suitability of the AP for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Cys24~Asp1 28. It is sometimes possible for the material contained within the vial of "Aprotinin (AP), Active Protein" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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