| Autor: |
Makhoul, Stephanie, Trabold, Katharina, Gambaryan, Stepan, Tenzer, Stefan, Pillitteri, Daniele, Walter, Ulrich, Jurk, Kerstin |
| Rok vydání: |
2019 |
| DOI: |
10.6084/m9.figshare.9830252 |
| Popis: |
Figure S1. Echicetin purification and qualitative validation. Echicetin was isolated from the snake venom of Echis carinatus sochureki by affinity chromatography followed by anion exchange chromatography using DEAE column. a Elution was performed with a gradient of NaCl, from 0 to 1 M at a flow rate of 1 ml/min. Two main peaks were separately eluted (P1 and P2) at around 62.5 and 167 mM, respectively. b Samples from P1 and P2 were analyzed by silver staining under non-reducing (NR) and reducing conditions (R) in a 15% SDS-gel. Bands detected under reducing conditions were cut from the gel, digested using trypsin, and analyzed by MS-analysis. The upper and lower bands of peak 1 were identified under the Uniprot IDs: P81017 (echicetin α-subunit) and P81996 (echicetin β-subunit), respectively. However, the upper band of peak 2 was detected as a sequence not related to Echis carinatus sochureki species, and the lower band was identified under the Uniprot ID: P81996 (echicetin β-subunit). For all experiments, echicetin from peak 1 was used. c Representative aggregation curves of human washed platelets (WP), which were stimulated under stirring conditions with EB or BSA-coated beads (as negative control). WP were pre-incubated with echicetin (EM) (25 μg/ml; 3 min) or with tirofiban (1.25 μg/ml; 1 min) prior to stimulation with EB. d Corresponding quantitative data of platelet aggregation expressed as maximum percentage of light transmission. Results are shown as means ± S.D. of 3 independent experiments with platelets from 3 healthy donors (****p |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
