Strategies to evade false positives in the in situ analysis of peptide antibiotics in SDS-PAGE gels.
In situ activity assay is one of the promising techniques for the characterization of peptide antibiotics. This assay was carried out for the peptide purified from a new bacterial isolate Paenibacillus alvei and commercial peptide antibiotic polymixin E. Towards this, the routine and new protocols were tried. Interestingly, the unexpected result of these experiments - the "switch over activity" has led us to have further investigations. Here, we have addressed the potential problem in the methodology of in situ assay and demonstrated a fool proof protocol to evade the false positive results.
Anandaraj
Balaiah
B
Centre for Biotechnology, Genetic Engineering Unit, Anna University, Chennai 600025, India.
Selvamanikandan
Athinarayanan
A
Mittal
Prabakar
P
Elavarasi
Natarajan
N
Thangadurai
Chinnathambi
C
Murugan
Vadivel
V
D000900:Anti-Bacterial Agents / Q000302:isolation & purification* / Q000494:pharmacology; D003091:Colistin / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005189:False Positive Reactions; D006090:Gram-Negative Bacteria / Q000187:drug effects; D008826:Microbial Sensitivity Tests / Q000379:methods*; D010455:Peptides / Q000302:isolation & purification*
D000900:Anti-Bacterial Agents; D010455:Peptides; D003091:Colistin