ELISA标准曲线制作方法.docVIP

ELISA标准曲线制作方法 一般而言，我们拟合ELISA标准曲线选用比较经典的Curve Expert 1.3或者Curve Expert 1.4软件。现在我们以Curve Expert 1.4为例，对ELISA标准曲线绘制方法进行详述。 Generally speaking, the standard fitting curve of ELISA is based on the classic software of Curve Expert 1.3 or Curve Expert 1.4. Now we will use the Curve Expert 1.4 to illustrate how to construct the ELISA standard curve. 1. 点击Curve Expert 1.4，打开应用程序，截图界面如下： Click Curve Expert 1.4, and then open the application programs. You will see the following screen shot. 2.在X轴输入标准品的OD值，Y轴输入相应的标准品的浓度，截图界面如下： Input the value of OD on the X-axis against the concentration of samples on the Y-axis. The following screen shot will appear like this. 3. 单击上图界面中的图标，出现如下界面 Click the icon showed on the above screen shot and you will see the following picture. 4. 单击上图界面中的ALL OFF 按钮，出现如下界面 Click the ‘All Off’ button , you can see the following screen shot. 5. 我们选择Sigmoidal Models这种曲线拟合模型进行ELISA标准曲线的拟合，点击OK，出现下图界面： We select the Sigmoidal Models to fit the ELISA standard curve. Click ‘OK’ ,and you will see the following screen shot. 注意：上图界面中的r值越高，即当r值越接近1时，所拟合的曲线就越好，所得到的数值可靠性也就也高。 Please note that the higher the r-value in the above picture ( or the closer the r-value is to 1), the more reliable values you will get. 6. 在上图空白界面，按[Ctrl]键+[L]键，出现如下界面 Please press the Ctrl plus L in the blank part of the above picture . The interface will appear like this. 在上图中输入相应的OD值（X值）后，点击Calculate即可计算出待测物的浓度（Y值），待测物稀释了多少倍就在所测值的基础上乘以稀释倍数即最终浓度。 After inputting the corresponding OD-value (x-value), click the button “Calculate”.The concentration (Y-value)of samples can be measured. The final concentration can be obtained by multiplying the the measured figure with the times the samples have been diluted. 7. 点击所绘制的曲线图中左上方按钮，可以得到如下ELISA曲线拟合方程 Click the button on the top left corner in the picture. Then you can get the ELISA curve fitting equation as follows. 8. 点击上图中 Copy 即可得到上图界面中的曲线方程数据 Logistic Model: y=a/(1+b*exp(-cx)) Coefficient Data: a = -