Standard Proteins (5) (QA-825)
Four kinds of protein standards were analyzed using IEC
QA-825
(a column for strong anion exchange chromatography). Since the pH value of the
eluent is 6.0 and the pI value of conalbumin is in the range of 6.0 to 6.8, the
conalbumin cannot be retained by the packing material and therefore it elutes
rapidly. The other proteins like transferrin, ovalbumin and trypsin are negatively
charged because their pIs are lower than 6.0 and therefore they are retained by
the packing material. These proteins are released from the surface of the packing
material and elute from the column by gradient elution with increasing salt concentration.
Sample :
1.
Conalbumin
2.
Transferrin
3.
Ovalbumin
4.
Trypsin inhibitor
Column : Shodex IEC QA-825 (8.0mmID*75mm)
Eluent : (A); 20mM Piperazine-HCl buffer(pH6.0)
(B); (A) + 0.5M NaCl
Linear gradient: 0min to 30min, 100% (A) to 50% (B)
Flow rate : 1.0mL/min
Detector : UV(280nm)
Column temp. : Room temp.