Analysis of Digested Protein using HPLC-UV
- Weight 1.0 to 2.0 mg each of insulin, BSA, monoclonal antibody, and thyroglobulin.
- Add 1.0 mL of SMART digest buffer and stir thoroughly.
- Place 200 µL of the above sample in a SMART digest tube and mix at a T=70°C, 1,400 RPM 2 hours using Thermomixer C.
- Centrifuge the prepared, mixed samples for injection.
The “SMART Digest kit” used in this research is a registered product of Thermo Fisher SCIENTIFIC.
The “Thermomixer C” used in this research is a registered product of Eppendorf.
HPLC Analytical Conditions:
|Column:||Develosil HSR C18 Peptide, 3 µm ( 4.6 x 75 mm)|
|Mobile Phase:||A) Water + 0.1% TFA
B) Acetonitrile +0.1% TFA
|Gradient:||Further details here|
|Flow Rate:||1.0 mL/min|
|Detection:||UV at 210 nm|
|Injection Volume:||(1) 10 µL
(2)-(4) 20 µL
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