Analysis of Digested Protein using HPLC-UV
Sample:
- Insulin
- BSA
- Mab
- Thyroglobulin
Protein Digestion:
- 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.
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HPLC Analytical Conditions: |
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|---|---|
| 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 |
| Temperature: | 50°C |
| Detection: | UV at 210 nm |
| Injection Volume: | (1) 10 µL (2)-(4) 20 µL |
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