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Cysteine-containing peptides were purified using an avidin column (Life Technologies/ Thermo Fisher Scientific).
Alkylated proteins were digested with trypsin and cysteine-containing peptides were captured using an avidin column (Life Technologies/Thermo Fisher Scientific).
To identify the enzyme responsible for b-VAD-FMK binding, we performed an affinity pull-down of the 'enzyme biotin-VAD-FMK' complex using an avidin column followed by protein separation on a 1D gel.
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After tryptic digestion, cysteine containing peptides were tagged with biotin, extracted using an avidin-coated column and identified by HPLC and mass spectrometry with collision-induced dissociation.
The labeled protein was further separated from any unlabeled protein by purifying biotin-labeled protein using a monomeric avidin column.
Immunohistochemistry was performed for freshly frozen sections by the biotin-avidin immunoperoxidase method using an avidin-biotin immunoperoxidase complex reagent (Vector Laboratories, Burlingame, CA, USA).
Immunoperoxidase stains were performed on de-waxed paraffin sections using an avidin-biotin horseradish peroxidase technique.
Immunohistochemical staining was performed using an avidin-biotinylated immunoperoxidase methodology.
Antibody detection was performed using an avidin-biotinylated enzyme.
All were visualized using an avidin-biotin detection system.
Twenty-eight consecutive colorectal adenocarcinomas were stained using an avidin-biotin indirect immunoperoxidase technique.
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