From Horse and Bovine Heart

Using the simple Reversed Phase HPLC Gradient Method shown in this note, it was possible to separate Horse and Bovine heart Cytochrome C. The Peaks were well separated and symmetrical. A linear detector response was observed over 2 orders of magnitude.

1. Cytochrome C from Horse heart
2. Cytochrome C from Bovine heart

Method Conditions
Column: Cogent Bidentate C8 300™, 5μm, 300Å
Catalog No.: 40008-75P-3M
Dimensions: 4.6 x 75mm
Mobile Phase:
A: DI Water / 0.1% Trifluoroacetic Acid (TFA)
B: Acetonitrile / 0.1% Trifluoroacetic Acid (TFA)

Time (minutes) %B
0 20
16 40
18 40
18.1 20

Post Time: 5 minutes
Flow rate: 0.5mL / minute
Detection: UV @ 214nm

Notes: Cytochrome C is used in the study of protein stability, folding, unfolding and molecular evolution. This protein is an efficient biological electron-transporter and is a universal catalyst of respiration.


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