英语翻译Measurement of OVA-specific antibodyOVA-specific IgG2a a

英语翻译
Measurement of OVA-specific antibody
OVA-specific IgG2a and IgG2b antibodies in serum were detected
by ELISA according to the method previously described by
Sjoelander et al.with some modifications (Sun & Liu,2008).In
brief,the wells of 96-well microtiter plates were coated with
100 ll of OVA solution (50 lg/ml in 50 mM carbonate buffer,pH
9.6) for 24 h at 4 \2C.The wells were washed 3 times with 200 ll
of PBS containing 0.05% (v/v) Tween 20 (PBS/Tween),and then
blocked with 200 ll of 5% FCS/PBS at 37 \2C for 1 h.After three
washings with PBST,100 ll of diluted serum samples (IgG2a,
1:100; IgG2b,1:50) or 0.5% FCS/PBS as control was added to triplicate
wells.The plates were then incubated for 1 h at 37 \2C,followed
by washing 3 times.Aliquots of 100 ll of horseradish
peroxidase-conjugated goat anti-mouse IgG2a or IgG2b (diluted
1:4000 in FCS/PBS) were added and incubated for 1 h at 37 \2C.After
washing,100 ll of TMB (3,30,5,50-tetramethylbenzidine) liquid
substrate was added to each well.The plate was incubated for
15 min at 37 \2C,and enzyme reaction was terminated by adding
50 ll/well of 2 N H2SO4.The absorbance was measured using the
ELISA reader at 490 nm with a 595 nm reference.Data were expressed
as the mean OD value of the samples minus the mean
OD value of the control.Results were expressed as log2 titers.
Where sets of serum samples have been subjected to within and
between group comparisons,ELISA assays of all the samples were
performed on the same day.
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测量卵子,特异性抗体
卵清蛋白特异性IgG2a和IgG2b抗体的血清进行检测
用ELISA法根据先前所描述的方法
Sjoelander等.与一些修改（星期日及刘,2008 ） .在
简言之,井96孔微量板涂层
100名当地雇员卵子解决方案（ 50 LG电子/毫升在50毫米的碳酸盐缓冲液,pH值
9月6日）为24小时4角的水井洗3次,200名当地雇员
的磷酸盐缓冲液含有0.05 ％ （ V / V ）的吐温20 （磷酸盐缓冲液/吐温） ,然后
封锁与200名当地雇员的5 ％的FCS /磷酸盐缓冲液在37 C的1小时经过三年
冲洗与PBST ,100名当地雇员稀释血清样品（ IgG2a ,
1:100 ; IgG2b ,1:50 ） ,或0.5 ％的FCS /磷酸盐缓冲液作为对照增加了三份
水井.板,然后孵育1小时37 ç ,其次
洗3次.
等分试样100镑辣根
过氧化物酶偶联羊抗小鼠IgG2a或IgG2b （稀释
1:4000功能界别/磷酸盐缓冲液）加和孵育1小时后在37角
洗衣机,100镑的四甲基联苯胺（ 3,30,5,50 -四甲基联苯胺）液体
底增加了每口井.该板孵育
15分钟在37 C和酶反应而终止增加
50 11 / 2不适用的以及硫酸.吸光度测定使用
酶联免疫吸附器在490纳米与595纳米参考.数据表示
作为平均OD值减去样本平均
OD值的控制.结果表示了log2滴度.
凡套血清样本受到内部和
组间比较,ELISA法检测的所有样本
表现在同一天.