Protein concentrations were determined using a spectrophotometric method (Iglesias et al., 2015 (link)). The homogeneity of the isolated protein was determined by SDS‐PAGE with a Mini‐Protean II mini‐gel device (Bio‐Rad; Milan, Italy), using 6% stacking (wt/vol) and 15% separation (wt/vol) gels under reducing conditions. Glycosylation analysis was performed on the gel after SDS‐PAGE using Pro‐Q Emerald 300 Glycoprot Probes Kombo (Life Technologies, Italy). The glycosylated proteins were visualized by the ChemiDoc™ XRS system. The amino terminal end of ledodin was sequenced by using automated Edman degradation performed on a Procise 494 sequencer (Applied Biosystems, Inc., Foster City, CA, USA) at the Service of Protein Chemistry at the Margarita Salas Center for Biological Research (CIBMS, Madrid, Spain).
Procise 494 sequencer
Manufactured by Thermo Fisher Scientific
Sourced in United States, Denmark, France
The Procise 494 sequencer is a laboratory instrument designed for DNA sequencing. It utilizes Sanger sequencing technology to determine the nucleotide sequence of DNA samples. The core function of the Procise 494 sequencer is to generate accurate and reliable DNA sequence data for scientific research and applications.
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Lab products found in correlation
3 protocols using procise 494 sequencer
1
Characterization of Recombinant Ledodin Protein
2
Histone H3 Cleavage Products Analysis
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A sample containing different APC-mediated cleavage products of histone H3 was transferred from an SDS-PAGE gel to a polyvinylidene difluoride membrane, stained with Coomassie Blue, and 2 bands were excised for N-terminal sequencing analysis. The analysis was performed by Alphalyse (Odense, Denmark) on an Applied Biosystems Procise 494 sequencer.
3
Edman Sequencing of Bacteriocin BacC1
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The N-terminal amino acids of the purified bacteriocin BacC1 were sequenced by Edman degradation using a Procise 494 sequencer (Applied Biosystems) equipped with a PTH-C18 column (Applied Biosystems, Roissy, France) for the quantitative determination of phenylthiohydantoin (PTH) amino acids.
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