Human plasminogen was isolated according to a published method (Deutsch and Mertz 1970) , with modifications as described elsewhere (Fleury and Angles-Cano 1991) , and was > 99% pure as assessed by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis (SDS-PAGE) and by amino-terminal sequence analysis. Urokinase was obtained from Microbix Biosystems (Mississauga, ON, Canada). Bovine serum albumin (BSA) was obtained from Sigma (St Louis, MO, USA), and aprotinin (Trasylol) was acquired from Bayer (Leverkusen, Germany).
Aprotinin trasylol
Manufactured by Bayer
Sourced in Brazil
Aprotinin (Trasylol) is a serine protease inhibitor used as a pharmaceutical product. It functions by inhibiting the activity of various proteolytic enzymes.
Automatically generated - may contain errors
Lab products found in correlation
Bovine thrombin, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Cm h2dcfda dcf, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Merck Group (1 mentions)
Fibrinogen, by Merck Group (1 mentions)
Ketamine, by Syntec (1 mentions)
Xylazine, by Syntec (1 mentions)
Soyabean trypsin inhibitor, by Merck Group (1 mentions)
Tumor necrosis factor α, by Thermo Fisher Scientific (1 mentions)
Hygromycin b, by Merck Group (1 mentions)
Plasminogen, by Enzyme Research (1 mentions)
Recombinant annexin 5, by Abcam (1 mentions)
Opti 4cn substrate kit, by Bio-Rad (1 mentions)
β actin sc 81178, by Santa Cruz Biotechnology (1 mentions)
S 2238 h d phe pip arg pna, by Werfen (1 mentions)
6 protocols using aprotinin trasylol
1
Purification of Human Plasminogen
2
Evaluating Anticoagulant Inhibitors: Protocols
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Human α-thrombin was purchased from CellSystems (St. Katharinen, Germany. rAPC (Xigris) was obtained from Eli Lilly (Indianapolis, USA). Argatroban (Argatra) was obtained from Mitsubishi Pharma (Düsseldorf, Germany). Aprotinin (Trasylol) was obtained from Bayer (Berlin, Germany). Recombinant hirudin (Refludan) was purchased from Pharmion (Hamburg, Germany). Biotinylated aptamers were synthesized and purified by Microsynth (Balgach, Switzerland). The fluorogenic peptide substrates Pefafluor TH (Thrombin, H-D-CHA-Ala-Arg-AMC) and Pefafluor PCa (APC, Pyr-Pro-Arg-AMC) were purchased from Loxo (Dossenheim, Germany).
3
Anticoagulant and Anti-inflammatory Activities of Vanadium (III) Chloride
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Vanadium (III) chloride (VCl3), sulfanilamide (SULF), N-(1-naphthyl) ethylenediamine dihydrochloride (NEDD), 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), purified coagulation factors (VII, IX, X, fibrinogen, plasminogen and kallikrein), Soya Bean trypsin Inhibitor (SBTI), Dulbecco’s modified Eagle’s medium (DMEM) and fetal calf serum (FCS) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). Aprotinin (Trasylol®) was obtained from Bayer do Brasil S.A. (São Paulo, Brazil). The chromogenic substrates for factor Xa (S2222, Bz-Ile-Glu-Gly-Arg-p-nitroanilide), plasma kallikrein (S2302, H-D-Pro-Phe-Arg-p-nitroanilide) and tissue kallikrein (S2366, H-D-Val-Leu-Arg-p-nitroanilide) were obtained from Chromogenix (Milan, Italy). CM-H2DCFDA (DCF), penicillin/streptomycin, EDTA and trypsin were from Life Technologies (Carlsbad, CA, USA). Ketamine and xylazine were obtained from Syntec (São Paulo, Brazil). Anti-lonomic serum was kindly donated by Butantan Institute (São Paulo, Brazil).
4
Venom-Induced Coagulation Disruption
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Lyophilized venom from adult specimens of B. jararaca snakes was obtained from the Laboratory of Herpetology, Butantan Institute. BjV was dissolved in sterile saline immediately before use. AEBSF, 1,10-phenanthroline (o-phe), bovine serum albumin (BSA), N-benzoyl-D,L-arginine-p-nitroanilide hydrochloride (BAPNA), and bovine thrombin were purchased from Sigma (USA), and sodium ethylenediamine tetraacetic acid (Na2-EDTA) from Bio-Rad (USA). Aprotinin (Trasylol) was obtained from Bayer (Brazil). To obtain rabbit anti-rat fibrinogen IgG, one rabbit was immunized i.m. with 500 µL of rat fibrinogen (4.32 mg/mL, [20] (link)) emulsified in 500 µL of Marcol-Montanide adjuvant; at fortnight intervals, the rabbit received four additional boosters in the same adjuvant. Anti-rat fibrinogen IgG was purified and biotinylated as previously described [21] . Rabbit anti-BjV serum was obtained as described elsewhere [22] (link). Rat thromboplastin was prepared as described elsewhere [23] ; briefly, dried thromboplastin was diluted in saline (40 mg/mL), maintained at 50°C for 20 min, and then refrigerated at 4°C overnight; the supernatant was used in clotting assays. All other reagents were of analytical grade or better.
5
Fluorometric Plasmin Activity Assay with Diverse Reagents
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Plasmin fluorescent substrate came from SensoLyte AFC Plasmin Activity Assay Kit *Fluorimetric* (Anaspec) and tPA Spectrofluor 444FL substrate was from American Diagnostica (ADF Biomedical, Neuville-sur-oise, France). Hygromycin B, bovine thrombin, carboxyfluorescein succinidimyl ester (CFSE), EDTA and lipopolysaccharide (LPS 0111:B4) were from Sigma-Aldrich (L'Isle d'Abeau, France). Bovine fibrinogen used for fibrin-agar zymography came from Bovogen Labs (Australia). Plasminogen used for plasmin substrate activity studies was from Enzyme Research Laboratories (ERL, UK). Sc-uPA, purified from human cell lines, was from Biopool AB (UK). The uPA standards for fibrin-agar zymographies were obtained using ACTOSOLV® (Eumedica). Lipofectamine 2000, fetal bovine sera were from Invitrogen (Cergy Pontoise). tPA (Actilyse) was from Boehringer-Ingelheim (Paris, France). Human PAI-1 (N-terminal cysteine, active fraction) came from Gentaur. Sheep antiserum raised against human tPA was prepared at the national institute for agronomic research (INRA, Clermont-Theix, France). Tumor Necrosis Factor-α was purchased from PeproTech (Rocky Hill, NJ). Recombinant annexin-V came from Abcam (France). Aprotinin (Trasylol) was a gift from Bayer HealthCare AG.
6
Measurement of Coagulation Factors
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Reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless stated otherwise. Antibodies for p38α MAPK: sc-535; p-p38α MAPK: sc-166182; NF-κB: sc-56735; β-actin: sc-81178, were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Opti-4CN substrate kit was acquired from Bio-Rad Laboratories (Hercules, CA, USA). Synthetic substrates for thrombin (S-2238™, H-D-Phe-Pip-Arg-pNa), factor XIa (S-2366, pyroGlu-Pro-Arg-pNa), plasma kallikrein and factor XIIa (S-2302™, H-D-Pro-Phe-Arg-pNA), factor Xa (S-2222™, Bz-Ile-Glu-Gly-Arg-pNa) and factor VIIa (SCP-0248, MeSO2-Cha-Abu-Arg-pNA)) were purchased from Chromogenix (Milan, Italy). Aprotinin (Trasylol®) was purchased from Bayer (São Paulo, Brazil). Ellagic acid, rabbit brain thromboplastin, phospholipids were purchased from Wiener Lab (Rosario, Argentina).
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