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yyc12596

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[交流] ELISA standards curve(excel做elisa的标准曲线)

from:
http://blog.sina.com.cn/s/blog_721358b20100ou1f.html

Step 1.

After Optical Densities (ODs) have been collected, enter each standard OD into a worksheet as shown.
  
Step 2.

Take the averages of the duplicate values either by adding them up and dividing by two on a calculator, or entering the formula =AVERAGE(A2+B2). Note: change the cell location from A2 and B2 to the desired cell location of each Standard OD.
  
Step 3.

Enter the Standard concentrations that each OD should represent next to their corresponding OD average.
   

Creating a Standard Curve

Step 1.

Highlight your data range and click on the "Chart Wizard" icon in the excel tool bar.
  
Step 2.

Choose XY (Scatter) and click "Next"
  
Step 3.

Click on "Series" located at the top of the Chart Wizard Window.

Make sure that your X Values are your concentrations and your Y values are your Average OD values.
Name your Series or add another Series if you want to compare Standard Curves
Click "Next"  
Step 4.

Add information to your chart such as titles and axis labels. Once you customize the chart to your preferences, click "Next".
  
Step 5.

Place your chart on a new Excel sheet or insert into your current worksheet.
  
Step 6.

Double click on the x-acis values and a "Format Axis" window will pop up. Click on the "Scale" tab on the top of the window. Check the "logarithmic scale" box.
  
Step 7.

Do the same to the y-axis
  
Step 8.

Highlight data points by clicking on one of them. Click on the "Chart" tab and on the top menu bar click on "Add Trendline".
  
Step 9.

A "Add Trendline" window will appear. Select Linear under "Trend/Regression type".
  
Step 10.

Go to the "Options" tab on the top of the window and Select "Display equation on chart" and "Display R-squared value on chart"
  
Step 11.

Click "OK" to view your chart with equations.



Analyzing Data

Now that you have your equation, you can solve for "x" and enter your sample OD values as Y. You can do this by using a calculator and manually solving for "X" or by inputing a formula as shown.

Use Excel Equation: y=0.062x+0.1579

Enter Formula: =(G7-0.1579)/0.0062)

where G7 is your OD value (select the appropriate cell for your data)
  

Note: Linear regression on Excel should only be used if software that can create a 4-Parameter (4-PL) logistic curve fit is unavailable. Please keep in mind that values that are represented at the low end of your curve will not be as reliable as values at the higher end of your curve.
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yyc12596

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EXCEL 行列互换
1、启动Excel,打开需要的工作簿,在当前工作表中,选中需要进行行列转换的单元格区域,如A1:X6。
  2、单击“编辑”菜单中的“复制”命令。
  3、单击要存放转置表区域的左上角单元格,如A7。
  4、单击“编辑”菜单中的“选择性粘贴”命令,系统“选择性粘贴”对话框,选中“转置”复选框。
  5、单击[确定]按钮,则行列转置后的表格出现在A7:F30单元格区域。现在把A1:X6单元格区域删除,则完成了表格的行列转置。
2楼2013-10-04 13:53:56
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yyc12596

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Standards  OD1          OD2
240020        182070
142680        144190
40980        48510
38400        33880
33100        29300
490650        25300
42500        42350
29240        32950

完全不靠谱的标准曲线数据。
3楼2013-10-08 09:12:06
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yyc12596

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Samples OD1        OD2
46250        40750
118710        109720
97410        78060
79630        65030
63880        51110
65860        52290
65160        56260
53900        48180
51560        43430

样本的浓度分析出来也只能做参考。不可用。因为标准曲线就没做好。
4楼2013-10-08 09:14:06
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yyc12596

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Standards            OD2        AVE-OD        OD-Blank        Concentrations
98XXXXX        98XXXXX        9855480        9577760        250
632XXXX        620XXXX        6264305        5986585        125
380XXXX        380XXXX        3804495        3526775        62.5
226XXXX        222XXXX        2243600        1965880        31.25
134XXXX        131XXXX        1329150        1051430        15.6
840XXX        810XXX        825535        547815        7.8
555XXX        567XXX        561300        283580        3.9
277XXX        278XXX        277720        0        0

这个是比较稳定的标准曲线值了。
5楼2013-10-08 09:24:40
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