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Team:METU Turkey/Results Discussion/CS
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<br>Loaded combined FPLC tubes from QS column(10/09/26) gave two close picks in CS column. First one corresponds to the CooA. The elutions of CooA are C13,C14 and C15. We combined them for further analyzes with EMSA and ITC and stored @+4. C16 AND C17 are combined to reload to CS column because they are contaminated another protein which is close to 40kDa. | <br>Loaded combined FPLC tubes from QS column(10/09/26) gave two close picks in CS column. First one corresponds to the CooA. The elutions of CooA are C13,C14 and C15. We combined them for further analyzes with EMSA and ITC and stored @+4. C16 AND C17 are combined to reload to CS column because they are contaminated another protein which is close to 40kDa. | ||
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<br>According to SDS-GEL, we combined 4,6,7,8 and 9 tubes containing 0,5ml and concentrated to 1ml for furhter analyzes such EMSA and ITF. We measured absorbance of combined @ 280,260 and 420 as respectively 2.76, 2.83 and 0.837. CooA concentration was determined as 7.75 uM according to extinction coefficient of heme(Ferric State) which is 108 mM-1 cm-1 according to Aono. We didnot take 5th tube because it contains much more impurities than other tubes. | <br>According to SDS-GEL, we combined 4,6,7,8 and 9 tubes containing 0,5ml and concentrated to 1ml for furhter analyzes such EMSA and ITF. We measured absorbance of combined @ 280,260 and 420 as respectively 2.76, 2.83 and 0.837. CooA concentration was determined as 7.75 uM according to extinction coefficient of heme(Ferric State) which is 108 mM-1 cm-1 according to Aono. We didnot take 5th tube because it contains much more impurities than other tubes. | ||
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Revision as of 23:04, 27 October 2010
10/09/26
Purification with CS BL21 pTriEx
Loaded combined FPLC tubes from QS column(10/09/26) gave two close picks in CS column. First one corresponds to the CooA. The elutions of CooA are C13,C14 and C15. We combined them for further analyzes with EMSA and ITC and stored @+4. C16 AND C17 are combined to reload to CS column because they are contaminated another protein which is close to 40kDa.
10/10/12 CS
Purification with CS BL21 pTriEx
According to SDS-GEL, we combined 4,6,7,8 and 9 tubes containing 0,5ml and concentrated to 1ml for furhter analyzes such EMSA and ITF. We measured absorbance of combined @ 280,260 and 420 as respectively 2.76, 2.83 and 0.837. CooA concentration was determined as 7.75 uM according to extinction coefficient of heme(Ferric State) which is 108 mM-1 cm-1 according to Aono. We didnot take 5th tube because it contains much more impurities than other tubes.
