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2 protocols using bovine seroalbumin

1

Analyzing Angiotensin Receptor Trafficking

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Fetal bovine serum, penicilin-streptomycin, Lipofectamine 2000, goat anti-mouse antibody coupled to Alexa 594 and Dulbecco’s modified Eagle’s medium (DMEM) were purchased from Invitrogen (Carlsbad, CA, USA). Bovine seroalbumin, paraformaldehyde, phosphate buffered saline (PBS) and the protease inhibitors cocktail were from Sigma Chemical Co. (St. Louis, MO, USA). Mouse anti Rab4, Rab11 or anti-CAV-1 antibodies were purchased from BD Biosciences. LysoTracker Red was from Molecular Probes. Ang-(1-7) was from Bachem. The plasmid containing MAS1R-YFP cDNA was obtained as previously described.17 (link) shRNA ß-arrestin2 was from SantaCruz. All other chemicals were analytical grade reagents of the highest purity available.
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2

Papain Purification from Carica papaya

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Papain (E.C. 3.4.22.2) from Carica papaya latex, crude powder (3.11 units/mg solid, one unit hydrolyzes 1.0 μmol of N α -benzoyl-arginine ethyl ester hydrochloride per min at pH 6.2 and 25 °C), dodecylamine and molecular sieves (4 Å) were obtained from Fluka. Bovine seroalbumin (BSA), Coomassie Brilliant Blue G-250, N α -benzoyl-arginine ethyl ester hydrochloride (Bz-Arg-OEt or BAEE), decylamine and N α -benzoyl-d,l-arginine p-nitroanilide hydrochloride (BAPNA) were from Sigma. Polyamide-6 (EP-700, particle size <800 μm, mean pore diameter 50-300 nm, specific surface area BET method 8.4 m 2 /g) was an Akzo (Obernburg, Germany) contribution. 1,4-Dithio-d,l-threitol (DTT) was purchased to Invitrogen (Carlsbad, CA, USA). Cetrimide was from Dishman Pharmaceuticals and Chemicals (India). Nutrient broth was obtained from Biokar Diagnostics (Beauvais, France). The rest of the chemicals and solvents used in this work were of analytical grade.
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