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| Peptide Synthesis | LPS-neutralizing activity of As-CATHs was determined using a ToxinSensorTM Chromogenic LAL Endotoxin Assay Kit (GenScript, Nanjing, China) according to the method described in our previous paper [30]. Briefly, LPS in the kit was dissolved with LAL water to 5 EU/ml. As-CATHs stock solutions (2 mg/ml) were prepared and serially diluted in a pyrogen-free 96-well microtiter plate (20 μl) (Polystyrene, Cat. No. 310109008, Fisher Scientific, USA). 20 μl LPS solution was added to each well, and the mixture was incubated at 37°C for 30 min to allow peptide binding to LPS. After that, LAL reagent, Chromogenic Substrate reagent, and Color Stabilizer #1, #2, #3 were successively added according to the kit instruction. The absorbance of the samples at 545 nm was measured. % LPS neutralization was calculated as (Ablank-Asample)/Ablank × 100, where Ablank represents the absorbance of blank control (20 μl LAL water + 20 μl LPS solution). | Get A Quote |
bstract Crocodilians are regarded as possessing a powerful immune system. However, the composition and action of the crocodilian immune system has remained unclear until now. Cathelicidins, the principal family of host defense peptides (HDPs), play pivotal roles in vertebrate immune defense against microbial invasions. However, cathelicidins from crocodilians have not been extensively studied to date. In the present study, six novel cathelicidins (As-CATH1~6) were identified and characterized from the endangered Chinese Alligator (Alligator sinensis). As-CATH1~6 exhibit no sequence similarity with any of the known cathelicidins. Structure analysis indicated that AsCATH1~3 adopt a random coil secondary conformat... More
