6040.2365赛默飞Viper 手拧接头系统Thermo

以非那西丁为探针采用高效液相色谱法快速测定细胞色素P450 1A2的酶活性

　　【摘要】  目的建立一种快速、高效的以非那西丁作为探针药物评价细胞色素P450 1A2（CYP1A2）酶活性的高效液相色谱-紫外检测方法。方法色谱柱为Agilent Zorbax SB-C18柱（50 mm×4.6 mm I.D.，5 μm）；流动相为乙腈-水（含0.01％七氟丁酸），梯度洗脱，流速2.0 ml・min-1；检测波长为244 nm。非那西丁与人CYP1A2酶在37 ℃温孵适当时间后，加入50 μl乙腈终止反应，10 000 g离心后取上清液进样分析测定。结果对乙酰

CAT ASSAY[TLC METHOD]

6. Spin down @ 14K RPM for 2-3'. 7. Resuspend in 100 µl 0.25M Tris pH 7.8 8. Freeze-thaw 3x (10'freeze-10'thaw). 9. Spin down debris @14K RPM for 5'. 10. Transfer 100 µl supernatant to a new set of tubes. 11. Set new set of tubes and add 40

Use of a Hemacytometer

as follows:  Cell concentration per milliliter = Total cell count in 4 squares x 2500 x dilution factor Example: If one counted 450 cells after diluting an aliquot of the cell suspension 1:10, the original cell concentration = 450 x 2500 x 10 = 11,250