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文献和实验min。②荧光素的准备 根据欲标记的蛋白质总量,按每毫克免疫球蛋白加0.01mg荧光色素,用分析天平准确称取所需的异硫氰酸荧光素粉末。也可用下述公式计算出免疫球蛋白、荧光素的量,还可以算出需加缓冲液的量。a.蛋白溶液:含量Amg/m1;容积Bml。b.总蛋白量(AXB)=Crag。c.C/20~C/10=Dmg(如蛋白含量低于20mg/ml,用C/10;如高于20mg/ml,用C/20)。d.荧光素FITC的量:(1/50~2/100)XC=Emg。e.巳0.5mol/L(pH9.5)碳酸盐缓冲
ELECTROPORATION OF ES CELLS AND ISOLATION OF H/R CLONES
, GIBCO-BRL 11811-031) need 150ug/ml active (however, each batch should be tested. Make up X100 in PBS and filter sterilize. NB. Each batch has number of ugs/mg active written on vial, you need to allow for this when making your calculation. FIAU
Dual- and Triple-Color Fluorospot
, in PBS containing 0.1% bovine serum albumin (BSA) (seeNote 8). Add 100 μl/well and leave the plate for 2 h at room temperature. c. Wash the plate as in step 6 above. d. Dilute secondary detection reagents, i.e., mAb anti-FITC
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