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By Cooper C., Packer N.
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Additional info for Amino Acid Analysis Protocols (Methods in Molecular Biology Vol 159)
This will weaken sample interaction with the detergent. 12. ProSorb cartridges should be prepared as directed by the manufacturer, using MilliQ water for all wash steps to reduce persistent background levels of contaminating amino acids. 13. Be careful with the manipulation of samples at every step. Be aware that proteins and peptides can irreversibly bind to glass and/or plastic tubes. If your sample is small, you can minimize losses in the cartridge by adding 100 µL of buffer to the prewetted membrane prior to the addition of sample.
And van Halbeek, H. (1993) Novel Asn-linked oligosaccharides terminating in GalNAcβ(1→4)[Fucα(1→3)]GlcNAcβ(1→•) are present in recombinant human Protein C expressed in human kidney 293 cells. Glycobiology 3, 597–608. 25. Chan, A. , Morris, H. , Eteinne, A. , Rogers, M. , et al. (1991) A novel sialylated N-acetylgalactosamine-containing oligosaccharide is the major complex-type structure present in Bowes melanoma tissue plasminogen activator. Glycobiology 1, 173–185. Purification of Proteins for AAA 31 3 Purification of Proteins Using UltraMacro Spin Columns or ProSorb Sample Preparation Cartridges for Amino Acid Analysis Li Zhang and Nancy Denslow 1.
1% TFA to reduce the concentration of ACN to less than 15%, prior to using the ProSorb cartidge. 10. Often proteins submitted for amino acid analysis contain detergents used in their preparation. Detergents can dramatically affect the binding of proteins to PVDF membranes. Table 1 summarizes the concentrations of detergents that are tolerable in a sample prior to binding to the membrane. Samples containing higher than these concentrations should be diluted to reduce the detergent and then added to the membrane in batches.
Amino Acid Analysis Protocols (Methods in Molecular Biology Vol 159) by Cooper C., Packer N.