| |
Click here to view
catalog description
|
CZE Application Sheet
Metallo-Proteins
- Short Analysis Time
- Simple, Easy, Fast
- Minimized Wall Adsorption
Method Conditions:
Voltage: 20kV
Capillary: MicroSolv CE100-SA Controlled EOF
50µm x 57cm, I=50cm
Injection: Hydrodynamic @ 0.5 psi
Run Buffers: 280mM Tricine w/30% MeOH
pH: 8.40
Detection: 200nm
Temperature: 15°C
|
Method:
Performed on a Beckman P/ACE 5000 instrument with liquid cooling, Metallothionein samples of rabbit liver
were purchased from Sigma Aldrich and prepared in ultra pure water at a concentration of 1.0mg/mL.
Injection was performed using pressure and Methanol was added to the run buffer to reduce EOF and further
minimize any wall adsorption. Capillaries were conditioned with run buffer for 30 minutes before flushing
the capillaries with water for five minutes. Polarity was normal.
Discussion and Rationale:
Separation at slight alkaline pH was chosen due to the natural state of these proteins where the metals are
still bound to molecules; at acidic pH the metals dissociate from the proteins. The CE100-SA Capillaries
exhibited near pH independence and very minimal wall adsorption especially when Methanol was added. The
E-gram shows rabbit liver MT-1 and MT-2 isoforms and sub-isoforms where all peaks are resolved. Difference
in the polymorphisms of the rabbit liver can be seen.
|
|
