Standard protein labeling procedure (lysine labeling):
- Dissolve the protein in an appropriate buffer* with pH 7.5-9.0 (e.g. PBS) at 1-10mg/mL concentration.
- Apply the appropriate amount of the stock solution of the reagent (1-5 molar eq. per lysine residue).
- Incubate at room temperature for 2 hours.
- If necessary, purify the protein-APN conjugate using size exclusion chromatography or ultrafiltration.
- The conjugate can be readily coupled with thiol-containing substrates by incubating the components in aqueous buffer (pH 6.5-9.0) at ambient temperature for 2 hours.
Standard protein labeling procedure (cysteine labeling):
Dissolve the protein in the appropriate buffer* with pH 6.5-9.0 (e.g. PBS) at 1-10mg/mL concentration.
Apply the appropriate amount of the stock solution of the APN-labeled molecule (1-5 molar eq. per free cysteine residue).
Incubate at room temperature for 2 hours.
If necessary, purify the protein conjugate using size exclusion chromatography or ultrafiltration.
*Note: avoid amine- and thiol-containing buffers.