CAJ | 학술논문

建立了一种基于125 I放射性标记并与Infinia Hawkeye 4 ECT检测相结合的检测蛋白分子位点密度的新方法。实验结果显示，P-选择素的位点密度与吸附浓度呈线性关系，且P-选择素在玻璃平面上的吸附能力高于在聚苯乙烯平面上的吸附。在相同P-选择素浓度下，对比标记 P-选择素和P-选择素单克隆抗体9 E1的放射性强度，结果发现，物理吸附作用下大概只有10％的 P-选择素是头部向上分布的。对比传统方法，新方法更为快速有效，且应用范围更广，新方法所测定的位点密度结果可以与流动腔实验相结合，从而获得一系列二维分子动力学参数。
건립료일충기우125 I방사성표기병여Infinia Hawkeye 4 ECT검측상결합적검측단백분자위점밀도적신방법。실험결과현시，P-선택소적위점밀도여흡부농도정선성관계，차P-선택소재파리평면상적흡부능력고우재취분을희평면상적흡부。재상동P-선택소농도하，대비표기 P-선택소화P-선택소단극륭항체9 E1적방사성강도，결과발현，물리흡부작용하대개지유10％적 P-선택소시두부향상분포적。대비전통방법，신방법경위쾌속유효，차응용범위경엄，신방법소측정적위점밀도결과가이여류동강실험상결합，종이획득일계렬이유분자동역학삼수。
Measuring the P-selectin site density on planar substrate is the primary event in understanding the mechanism of P-selectin-induced cell adhesion on molecular level through the flow chamber experiment . However ,the sensibility of fluorescence labeling fails to detect the small amount of P-selectin ;and the traditional gamma counter cannot detect the radiation on planar surface although the sensibility of 125 I labeling is enough .Thus ,a new method to detect the density of 125 I labeled protein on planar surface is established by using Infinia Hawkeye 4 ECT . The results illustrate that the P-selectin site density is linear with their concentration ,and the glass planar surface shows better adsorption than polystyrene for P-selectin .For the same P-selectin concentration ,the difference in radiation intensity between direct P-selectin with 125 I and its antibody 9E1 with 125 I label bound on P-selectin suggests that only about 10% coated P-selectins are in its head up position .Comparing with the traditional method ,this method is more efficient and can be applied more broadly .Finally ,the results measured by using this new method can be combined with the flow chamber experiment to obtain a series of 2D molecular dynamic parameters .