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# Generating an ELISA standard curve in Excel

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MD Bioproducts Resource: Generating an ELISA standard curve in Excel:
If software capable of generating a 4-Parameter Logistic curve fit is unavailable, a standard curve using linear regression analysis in excel can be used.

Entering Data into Microsoft Excel

 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

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.

Sample Data comparing standard curve values using 4-PL vs Linear regression.

 Concentration (ng/mL) Mean OD 4-PL (ng/mL) Linear regression (ng/mL) 300 1.996 300.05 296.44 150 1.153 150.42 160.47 75 0.649 74.33 79.18 37.5 0.392 37.46 37.73 18.75 0.268 19.72 17.73 9.38 0.198 9.26 6.44 4.69 0.165 4.15 1.11

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Posted By: MD Bioproducts on 1/14/2010 10:53:20 AM

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