 | Acta Crystallographica Section F Acta Crystallographica Section F STRUCTURAL BIOLOGY COMMUNICATIONS |
journal menu
![[Figure 1]](en5068fig1mag.jpg)
|
|
Figure 1
Purification of agkicetin-C. (a) 1 g crude venom from D. acutus was dissolved in 20 ml buffer A (20 mM Tris–HCl pH 8.0) and centrifuged at 12 000 rev min−1 for 15 min. The supernatant was applied onto a DEAE-Sepharose Fast Flow column (1.6 × 40 cm) pre-equilibrated with buffer A. The column was washed at a flow rate of 120 ml h−1 initially with buffer A until peaks 1 and 2 were eluted and then with an 800 ml linear NaCl gradient (0–200 mM in buffer A). Finally, the column was exhaustively eluted with a high concentration of NaCl (500 mM in buffer A). Peak 4 containing agkicetin-C was pooled. (b) Peak 4 mentioned above was condensed and dialyzed against buffer B (50 mM sodium citrate pH 5.0) overnight at 277 K and then applied onto a CM-Sepharose Fast Flow column (1.6 × 40 cm) pre-equilibrated with buffer B. A similar strategy to the linear NaCl gradient used above was applied with buffer B and peak 2 was pooled. (c) Peak 2 mentioned above was condensed and dialyzed against buffer A overnight at 277 K and then applied onto a Mono Q HR 5/5 column (3.6 × 100 mm) pre-equilibrated with buffer A. The column was washed at a flow rate of 60 ml h−1 initially with 5 ml buffer A and then with a linear NaCl gradient (0–200 mM in buffer A). Finally, the column was eluted with a high concentration of NaCl (500 mM in buffer A) as mentioned above. The main fraction collected proved to be agkicetin-C. |
![[Figure 1]](en5068fig1.jpg)
| STRUCTURAL BIOLOGY COMMUNICATIONS |
ISSN: 2053-230X
