现代食品科技2024，Vol.40Issue(4)：296-302,7.DOI:10.13982/j.mfst.1673-9078.2024.4.0968
乳及乳制品中乳铁蛋白含量的快速SDS-PAGE荧光检测方法
Rapid SDS-PAGE Fluorescent Detection Method for Lactoferrin Content in Milk and Dairy Products
摘要
Abstract
A novel dye(Chromeo P503,P503)was used as an alternative to Coomassie brilliant blue to address the drawbacks associated with the laborious and time consuming(≥5 h)staining and destaining processes.This facilitated the establishment of a rapid and simple sodium dodecyl sulfonate-polyacrylamide gel electrophoresis(SDS-PAGE)method for the detection of lactoferrin in milk and dairy products.P503 dye-labeled protein emits strong red fluorescence,which could be completed in one step within 30 min.The interference of lactoferrin iron saturation on fluorescence signal was eliminated after apolactoferrin preparation to ensure the accuracy of quantification.Qualitative and quantitative detection of lactoferrin was achieved based on the presence of a 78 ku protein band and its gray value in the gel image.Under optimized experimental conditions,the linear range was 15～75 μg/mL,and the limit of detection was 5.11 μg/mL,with a recovery of 75.79%～108.09%,indicating that this method has high sensitivity and accuracy.The combination of the SDS-PAGE and P503 methods enabled the rapid,simple,and accurate detection of lactoferrin in milk and dairy products,eliminating the need pretreatment of the heparin affinity column.P503 dye labeling can facilitate rapid and low-cost quantitative detection of other milk proteins using SDS-PAGE,providing a valuable tool for dairy quality control,nutrition evaluation,and processing technology research.关键词
乳铁蛋白/十二烷基硫酸钠-聚丙烯酰胺凝胶电泳/Chromeo P503/快速标记Key words
lactoferrin/sodium dodecyl sulfate-polyacrylamide gel electrophoresis/chromeo p503/rapid staining引用本文复制引用
李汉芳,黄珍,路梦凡,孙娜娜,徐丹,徐秦峰..乳及乳制品中乳铁蛋白含量的快速SDS-PAGE荧光检测方法[J].现代食品科技,2024,40(4):296-302,7.基金项目
陕西省教育厅项目(22JC008) （22JC008）
陕西省科技厅项目(2021NY-174) （2021NY-174）
