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Measurement of SNAREs:lipids ratio

YF Yuhui Fu
BD Binbin Ding
XL Xiaoxia Liu
SZ Shangang Zhao
FC Fang Chen
LL Linsen Li
YZ Yi Zhu
JZ Jingxuan Zhao
ZY Zhen Yuan
YS Yafeng Shen
CY Chaofeng Yang
MS Mengle Shao
SC She Chen
PB Perry E. Bickel
QZ Qing Zhong
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We used the Stewart Method to measure the phospholipid concentration of adiposomes. To avoid fluorescence dye to interrupt the 472 nm absorbance reading, we made adiposomes as how we made the adiposomes in FRET assay except that there are no fluorescence dye included. On the other side, we made a standard curve by liposomes with the same phospholipid compositions (66.6% DOPC, 33.3% POPE), to get a function of absorbance reading at 472 nm in terms of phospholipid amount. Based on this function and the 472 nm reading of the adiposomes we freshly made, we can get its phospholipid amount. Furthermore, the protein amounts of the adiposome samples were estimated by comparing the corresponding SNARE protein bands with BSA bands on SDS-PAGE. Then the ratio between phospholipids and proteins can be calculated.

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