Standard laboratory chemicals and protease inhibitor cocktail (PIC) were purchased from Sigma-Aldrich (St. Louis, MO, USA) and Research Products International (Mt. Prospect, IL, USA). Q-sepharose, Heparin and S200 size exclusion chromatography resins were from GE Healthcare (Pittsburgh, PA, USA). Ni2+-NTA agarose was from Gold Biotechnology (St. Louis, MO). Biogel-P4 resin was from Bio-Rad Laboratories (Hercules, CA, USA). Enzymes for molecular Biology were from New England Biolabs (Ipswich, MA, USA). Oligonucleotides were purchased from Integrated DNA Technologies (Coralville, IA). Fmoc-4-amino-phenylalanine was from Angene International Ltd. (Nanjing, China). Commercial 4-azidophenylalanine was purchased from Chem-Impex International (Wood Dale, IL, USA). MB543 DBCO was purchased from Click Chemistry Tools (Scottsdale, AZ, USA). Alexa Fluor 594 DIBO alkyne and BL21Ai cells were purchased from ThermoFisher Scientific (Waltham, MA, USA).
Biogel p4 resin
Manufactured by Bio-Rad
Sourced in United States
Biogel-P4 resin is a size exclusion chromatography media used for the separation and purification of macromolecules, such as proteins, nucleic acids, and polysaccharides, based on their molecular size and shape. The resin is composed of cross-linked polyacrylamide beads and is suitable for a range of applications, including desalting, buffer exchange, and molecular weight determination.
Automatically generated - may contain errors
Lab products found in correlation
Q sepharose, by GE Healthcare (1 mentions)
Heparin, by GE Healthcare (1 mentions)
S200 size exclusion chromatography, by GE Healthcare (1 mentions)
Bl21 ai cells, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Enzymes for molecular biology, by New England Biolabs (1 mentions)
Oligonucleotide, by Integrated DNA Technologies (1 mentions)
α1 2 3 mannosidase, by New England Biolabs (1 mentions)
α1 6 mannosidase, by New England Biolabs (1 mentions)
Nh4ch3coo, by Merck Group (1 mentions)
2 protocols using biogel p4 resin
1
Protein Purification Using Chromatography
2
Fractionation and Characterization of Oligosaccharides
Check if the same lab product or an alternative is used in the 5 most similar protocols
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A 0.5 × 20 cm column was packaged with 6 ml of the BioGel P4 resin (Bio-Rad) and conditioned with a 25 mM NH4CH3COO (Sigma–Aldrich) buffer. Ten nanomoles of the total fOS were loaded into the column and 24 fractions (500 ul per fraction) were collected. Sample were dried in vacuum and later resuspended in water and desalted by ion exchange chromatography. Ten percent of each fraction was collected for FACE analysis, and the remaining 90% was divided in half and used for macrophage stimulation in biological duplicates. For the mannosidase digestion, 10 nmol of fOS or the synthetic ManGlcNAc disaccharide were treated with 40 U of α1-6 mannosidase (New England Biolabs) and 32 U of α1-2, 3 mannosidase (New England Biolabs). Samples were then mixed with 3 volumes of cold ethanol and centrifuged at 20,000 × g for 20 min at 4 °C. The supernatant was dried in vacuum and later resuspended in water and desalted by ion exchange chromatography. Ten percent of the digested product was collected for FACE analysis and the remaining 90% was divided in half and used for macrophage stimulation in biological duplicates.
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