Download e-book for kindle: Antimicrobial Peptides: Methods and Protocols by Paul R. Hansen (eds.)
By Paul R. Hansen (eds.)
This quantity offers uncomplicated and complicated laboratory protocols for the non-specialist and the skilled researcher. Chapters are divided into 3 elements overlaying synthesis and characterization of AMPs, learning the interplay of AMPs with version structures or micro organism, and assaying chosen organic actions of AMPs. Written within the hugely winning Methods in Molecular Biology series layout, chapters comprise introductions to their respective subject matters, lists of the mandatory fabrics and reagents, step by step, without problems reproducible laboratory protocols, and tips about troubleshooting and fending off identified pitfalls.
Authoritative and state of the art, Antimicrobial Peptides: tools and Protocols aims to make sure winning leads to the additional research of this important field.
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Additional info for Antimicrobial Peptides: Methods and Protocols
This method can be used either to design AMPs de novo or suggest mutations to existing AMP sequences that might increase their selectivity [7–9]. The design of AMPs in any case necessitates knowledge-based decisions that in turn require a sound prior understanding of how structural and physical attributes of this class of peptides affect their mechanism of action against bacteria (mainly, but not only, membrane effects) and cytotoxic activity against host cells. In this respect, peptides of the amphipathic α-helical structural group are by far the most studied and best understood among AMPs, and within this group, those of anuran origin are preponderant.
2, steps 1–3) and then repeat steps 1–4. Remove the piston and place the syringe on a suction plate. 2. If any resin residue is left on the pistonhead, wash it down with DMF. Wash with DMF (3×), DCM (3×), and DMF again (5×). Transfer 2–3 mL of the 20 % piperidine in DMF solution into the syringe. 2). Repeat steps 1 and 2 twice. For peptides longer than 10 residues, we recommend extending the time of the last two deprotection cycles to 7 min each. Put the piston back in place and change the syringe tip.
Aureus, MIC data is significantly less abundant). Always verify that a comparable bacterial load (CFU/ml) and similar medium conditions were used to determine the MIC (see Note 1). 3. Cytotoxic activity is assessed in terms of HC50, as the hemolysis assay is by far the most frequent assay used for AMPs. Even so, it is the limiting data item, as it is tenfold less abundant than E. coli MIC values. Always verify that comparable cell loads and conditions were used (see Note 2). 4. The ratio of hemolytic to antibacterial activity provides a commonly used selectivity index, SI = HC50/MIC (see Note 3).
Antimicrobial Peptides: Methods and Protocols by Paul R. Hansen (eds.)