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2 protocols using phycoerythrin pe antihuman cd41a antibody

1

Platelet Activation and Signaling Assay

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Hepes, apyrase, adrenaline [(−)-epinephrine (+)-bitartrate salt)], acetylsalicylic acid (ASA), bovine serum albumin (BSA), human fibrinogen, PGE1, LY 294002, wortmannin, digitonin, nigericin, EIPA, BCECF-AM, acetazolamide, indometacine, phentolamine, TRIS (tris(hydroxymethyl)aminomethane), DIDS, Ponceau Red, and TBST buffer—Millipore, Tirofiban (Aggrastat) were acquired from Sigma (Merck KGAA, Darmstadt, Germany). Type I collagen was acquired from Chrono-log. Phycoerythrin (PE)-antihuman CD41a antibody, FITC-antihuman CD62P (P-selectin), and FITC-annexin V were acquired from Becton Dickinson (Franklin Lakes, NJ, United States). Rauwolscine hydrochloride, PSB 0739, KB-R7943 mesylate, 2-APB, MRS 2500, SQ 22536, and BRL 44408 maleate were acquired from Tocris. BAPTA-AM was acquired from Abcam (Cambridge, UK). Fura-2-AM and AlexaFluor(AF)647-annexin V were acquired from Thermo Fisher Scientific (Waltham, MA, United States). Bovine thrombin was acquired from was acquired from Synthaverse S.A., Lublin, Poland. Other chemicals were acquired from Sigma (Merck).
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2

Platelet Function Assays with Inhibitors

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Hepes, apyrase, adrenaline [(-)-epinephrine (+)-bitartrate salt)], ADP disodium salt, human fibrinogen, acetylsalicic acid (ASA), bovine serum albumin (BSA) were from Sigma (Merck). Type I collagen was from Chrono-log. Batroxobin (Dade Behring) and tissue factor (Innovin) were from Dade Behring. DiO (3-octadecyl-2-[3-(3-octadecyl-2(3H)-benzoxazolylidene)-1 propenyl]-, perchlorate, AlexaFluor (AF)488-human fibrinogen and AF647-annexin V were from Thermo Fisher. Phycoerythrin (PE)-anti human CD41a antibody and FITC-annexin V were from Becton Dickinson. Recombinant tissue plasminogen activator (Actilyse, Boehringer Ingelheim). Rauwolscine hydrochloride and PSB 0739 were from Tocris. Tirofiban (Aggrastat) was from Merck. Nadroparinum calcinum (Fraxiparine) was from Glaxo Smith Kline). PPACK hydrochloride was from Santa Cruz Biotech. Other chemicals were from Sigma (Merck). Throughout the text “rauwolscine” means a water solution of Rauwolscine hydrochloride.
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