(1) Assay Card

General Information
Summary Kalata B1 showed concentration dependent erythrocyte lysis.
Condition Sheep erythrocytes were washed six times with a 10-fold excess of PBS by serial centrifugation at 4000 rpm, then a 1% (v/v) solution of red blood cells in PBS was prepared. Test peptides were added to 20 ul of 1% red blood cell solution to make a total sample volume of 100 ul and final peptide concentrations of 50, 100 and 200 uM. The samples were incubated for one hour at room temperature, centrifuged and the absorbance at 415 nm of 20 ul of supernatant in 600 ul water was measured. Controls of 1% SDS for complete haemolysis and PBS for zero haemolysis were run concurrently.
Result Haemolytic assays showed a concentration dependent erythrocyte lysis for the wild-type cyclic kalata B1. Sheep blood was found to be more resilient than human blood, with a maximum of 14% haemolysis observed at 200 lM peptide concentration. In contrast with the wild type peptide, all acyclic permutants had no observable haemolytic activity up to 200 uM.
AssayType Hemolytic

Simonsen SM, Daly NL, Craik DJ (2004) Capped acyclic permutants of the circular protein kalata B1. FEBS Lett 577:399-402

Proteins Assayed N-KB1-C