Analysis Tools
ELISA (Immunometric/Sandwich) Analysis Tools
These resources are provided to serve the analysis needs of researchers using our immunometric/sandwich ELISA kits.
If you have questions on the use of our tools, or feel something is missing, please contact Technical Services.
The Workbooks
Currently, Cayman Chemical uses two Excel workbooks to perform immunometric analysis. Both are available for your download and use.
Download
Your downloading these files implies acceptance of the license agreement listed below and present in each workbook.
- ImmunoTriple~ 600 Kb XLS formatted for assays performed in triplicate
- ImmunoDouble~ 600 Kb XLS formatted for assays performed in duplicate
Trouble downloading? Right-click the link and select "Save Target As..."
These instructions are repeated on the first sheet of each workbook.
ImmunoTriple Worksheet
ImmunoDouble Worksheet
Definition of Key Terms
| Term | Definition |
|---|---|
| Blank | Background absorbance caused by Ellman's Reagent |
| Std0# (Standards) | Standards #1 through #8 made by serially diluting Std01 seven times. See diagram (concentrations may vary depending on the assay) Standards serial dilution |
| Blank | Background absorbance caused by Ellman's Reagent |
| Std0# (Standards) | Standards #1 through #8 made by serially diluting Std01 seven times. See diagram (concentrations may vary depending on the assay) Standards serial dilution |
| Smpl# (1:#) | Samples and their associated dilution factors [Dilutions must be entered in the format shown] |
| Ctrl# (1:#) | Positive controls and their dilutions (Cayman Chemical uses 1:5 dilutions of Std01 and Std03) |
Workbook Sheets
Analysis
| Field | Explanation |
|---|---|
| LAYOUT | Contains sample names and associated dilution factors |
| RAW DATA | Data entered into the worksheet in the format defined by the LAYOUT section |
| SUBTRACTED | The AvgRawBlnk is subtracted from each cell |
| CONC | The concentration for each cell is calculated based on the absorbance value, constants from the linear regression curve (see StdCurve section), and the dilution factor from the LAYOUT section |
| STANDARDS | The Avg and Recalc values are calculated for each concentration value. Recalc are the standard concentration values calculated from the regression constants |
| Samples | This table is compiled from the preceding sections. It includes
|
| STATS | y-int and slope are obtained from the linear regression fit of the calibration data. |
| FILENAME | The filename given to the Excel workbook. |
StdCurve
This is the calibration curve of the concentration standards plotted versus their absorbance values. The data are fit to a linear regression line with the general equation: y = m x + b.
Instructions
The worksheets were designed to be as "hands-off" as possible. The following instructions assume you have loaded your plate according to one of the plate layouts shown above, and have the data in a form that can be imported (or pasted) directly into a spreadsheet. To analyze your data:
- Select your assay from the pull-down, scrollable menu.
- Input your sample names into the appropriate cells
- Input the sample dilutions into the appropriate cells in the format shown (e.g., 1:5)
- Paste/type your data into the RAW DATA section (starting in cell B14) of the Analysis worksheet. Select the "Values" option in the "Paste Special..." dialog box to maintain formatting
All concentration values, and relevant statistics, are automatically calculated. The Samples table is then filled in with the data. All averages and statistics are calculated based on all replicates for each sample. It is the responsibility of the individual researcher to decide if any data can be eliminated. Caution should be exercised, however, when doing this! Cayman Chemical has examined statistical methods to test for the presence of outliers in immunometric assays. We find that individual replicates are rarely flagged as outliers, and cannot be eliminated from the experiment.
Note The sole purpose of the data contained in this workbook is to demonstrate the functions of the individual spreadsheets.
Copyright and Disclaimer
Copyright © 2026 Cayman Chemical Company, Inc. All rights reserved.
Redistribution and use in source and binary forms, with or without modification, are permitted provided that the following conditions are met:
- Redistributions of source code must retain the above copyright notice, this list of conditions and the following disclaimer.
- Redistributions in binary form must reproduce the above copyright notice, this list of conditions and the following disclaimer in the documentation and/or other materials provided with the distribution.
THIS SOFTWARE IS PROVIDED BY CAYMAN CHEMICAL COMPANY, INC. "AS IS" AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL CAYMAN CHEMICAL BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED OF THE POSSIBILITY OF SUCH DAMAGE.
The views and conclusions contained in the software and documentation are those of the authors and should not be interpreted as representing official policies, either expressed or implied, of Cayman Chemical Company, Inc.