For fluorescent labeling of PF201, PF201 was dissolved in 0.1 M NaHCO3 and then mixed with 100 mg/mL solution of Cyanine3 NHS ester (Cy3-NHS ester; Lumiprobe) at a ratio of 9:1. Following 4-h reaction under dark conditions, the Cy3-labeled PF201 solution was desalted using a Sephadex G-10 gel filtration column (GE Healthcare). The Cy3-labeled PF201 (100 mg/kg) was orally administered to 7-week-old male ddy mice (Shimizu Laboratory Supplies Co., Ltd.), and the stomach, small intestine, portal vein, and liver were collected at 1 h after the administration. The organs were homogenized in phosphate buffered saline (PBS). Extracts obtained from each organ were analyzed by HPLC (Nexera XR; Shimadzu) to detect Cy3-labeled peptides.
Sephadex g 10 gel filtration column
Manufactured by GE Healthcare
Sephadex G-10 is a gel filtration column used for the separation and purification of small molecules and peptides. It is composed of cross-linked dextran beads that act as a molecular sieve, allowing the separation of substances based on their molecular size and weight. The column is designed to effectively separate compounds with molecular weights up to 700 daltons.
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Lab products found in correlation
2 protocols using sephadex g 10 gel filtration column
1
Fluorescent Labeling and In Vivo Tracking of PF201
2
Enzyme Digestion and Peptide Characterization
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The inactivated
enzyme was digested overnight at 37 °C with trypsin and chymotrypsin
(enzyme: trypsin/chymotrypsin—50:1 w/w) followed by separation
with a Sephadex G-10 gel filtration column (GE Healthcare Life Sciences).
The peptide pool was characterized by Q-TOF ESI XENO XS mass spectrometry
to identify the molecular weights of the digested separated fragments
(abbreviated here as EDPs, enzyme-digested peptides) as well as to
identify any residual undigested enzyme.
enzyme was digested overnight at 37 °C with trypsin and chymotrypsin
(enzyme: trypsin/chymotrypsin—50:1 w/w) followed by separation
with a Sephadex G-10 gel filtration column (GE Healthcare Life Sciences).
The peptide pool was characterized by Q-TOF ESI XENO XS mass spectrometry
to identify the molecular weights of the digested separated fragments
(abbreviated here as EDPs, enzyme-digested peptides) as well as to
identify any residual undigested enzyme.
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