GainData® ELISA Analysis Tool
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Yes, GainData® is completely free for registered members. To get started, please visit the arigo Member Center to create a free account. Once signed in, you will have immediate and full access to all GainData® features.
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No, there is no limit. GainData® allows you to analyze data as many times as needed. If the "Analyze" button becomes unresponsive during consecutive runs, please check the following:
1. Ensure all 96 wells are completely filled. Do not leave any well blank (enter 0 for empty or unused wells).
2. Check your raw data for non-numeric characters (such as commas ,, colons :, or scientific notation like E+n).
3. If the formatting is correct but the issue persists, press Ctrl + F5 to force-refresh your browser.
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．You can rest assured that your results are highly accurate. GainData® was independently developed in-house as a dedicated ELISA analysis tool. Our calculation models are rigorously validated and yield results highly consistent with mainstream statistical software like GraphPad Prism.．Furthermore, the accuracy and reliability of GainData® are well-recognized in the scientific community, having been cited in top-tier journals, including:。Nature Medicine (PMID: [35668177])。Nature (PMID: [35197629])
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This is almost always caused by data formatting errors. Please verify the following:
1. Decimal Format: The tool only accepts standard numeric formats. Ensure periods . are used as decimals. Replace any commas , with periods ., and remove any dashes -.
2. No Blank Fields: Every well in the grid must contain a value. Fill any unused or blank wells with 0.
3. No Scientific Notation: Do not use scientific notation (e.g., E+n). Convert these into standard numbers.
4. Sample ID Limit: Ensure that your total number of Sample IDs does not exceed 96.
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GainData® is strictly structured around the standard 96-well plate format (8 rows × 12 columns). Your data must be organized into this exact 8×12 grid in Excel before copying and pasting. Even if you only used a few wells, the rest of the grid must be filled with 0 to maintain the layout.
For a clear demonstration, click the "Demo data" button on the tool page or watch our Data Import Video Tutorial.
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If your data is purely numeric and properly formatted but still won't paste, try reloading the webpage. When making manual adjustments directly within the "Data Importing" box, click on the specific character and use the Delete or Backspace key on your keyboard.
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We highly recommend pasting the Raw OD data directly from Excel. GainData® features an automated background correction option. By simply selecting "Subtract minimum OD value" under the method settings, the tool will automatically handle the blank subtraction for you.
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Yes. CCK-8 data can be directly pasted and analyzed. For a standard BCA protein assay, choosing the Linear regression model is generally sufficient to achieve accurate quantification. However, we always recommend checking the specific curve-fitting guidelines provided by your assay kit supplier.
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These options apply different mathematical treatments to your data:
．Set Blank as Standard Data: The software treats the blank well as a standard point with a concentration of 0 and includes it in the standard curve calculation.
．Subtract minimum OD value:
。If blank wells are defined, the software subtracts the blank OD value from all other wells to eliminate background noise.
。If no blank wells are defined, it subtracts the minimum OD value found among the standards.
For most standard ELISA data processing, we recommend selecting both options to ensure the highest accuracy.
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These functions serve as an auto-fill shortcut when entering standard concentrations that have been serially diluted. Once you enter the concentration of your first standard, you can use these buttons to quickly populate the rest. This setting is strictly for defining the standard curve concentrations and does not apply a dilution factor to your unknown samples.
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If no dilution was performed, you do not need to change anything; leave it at the default value of 1. If your samples were diluted, please refer to our Dilution Factor Calculation Video.
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In the Four-Parameter Logistic (4PL) model, "d" represents the Upper Asymptote (the maximum theoretical response), which is normally a non-zero positive value. If you see d = 0, it typically indicates that the data was heavily normalized prior to import, or that a non-standard, simplified curve model is being forced.
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"cal.conc" stands for Calculated Concentration. It is the final concentration value derived by mapping your sample's raw OD value against the generated standard curve.
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Polynomial (quadratic) regression is a second-order polynomial model, which uses a completely different mathematical approach compared to the Four-Parameter Logistic (4PL) model. Because ELISA binding responses level off at higher concentrations due to saturation, the 4PL model best reflects the true biological characteristics of ELISA data.
For a detailed technical explanation, please refer to this reference paper: ScienceDirect - ELISA Data Analysis via 4PL.
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Generally speaking, a coefficient of determination R² value of greater than or equal to 0.95 is considered acceptable for biological assays. The 4PL curve-fitting model optimized by GainData® routinely provides excellent, highly trustworthy R² values for ELISA datasets.
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GainData® rounds the R² value from the fourth decimal place to the third decimal place. Because the 4PL regression model fits ELISA data exceptionally well, the R² value is often extremely close to 1 (e.g., 0.9996). When rounded to the third decimal place, it displays as 1 or 1.000.
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This is a natural characteristic of log scaling. A log scale line does not use standard base-10 even subdivisions; instead, it utilizes roughly 0.17 subdivisions based on logarithmic pacing, which creates a visible shift compared to linear grids.
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Currently, GainData® does not automatically calculate CV% values. However, you can easily compute it manually using the Standard Deviation (SD) and Mean provided in your report with the following formula:
CV = Standard Deviation (SD) /Mean
Alternatively, we highly recommend exporting your results in Excel format and using standard Excel formulas (=STDEV() / Average) to batch-calculate CV% values quickly.
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The primary purpose of the GainData® calculator is to convert raw absorbance (OD) values into concentration values. If you have already mathematically calculated the log of the concentration, the data transformation is already complete, and you do not need to feed it through the ELISA calculator.
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"Out of Range" indicates that a sample's OD value falls outside the upper or lower limits of your standard curve (above the upper asymptote d, or below the lower asymptote a). This means the sample concentration cannot be accurately read within the current linear range of the assay.
【Reporting & Corrective Actions】
．Above the Highest Standard: The sample is too concentrated. You must dilute the sample and re-test it, then multiply the result by the dilution factor. Alternatively, you can note it as > [Highest Standard Concentration] ng/ml.
．Below the Lowest Standard: The sample concentration is too low. You can choose to report it as ND (Not-Detected), or concentrate your sample and run the assay again.
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1. Mathematical Constraints: Logarithmic scales cannot process a value of 0 or negative numbers. If background subtraction results in any well value becoming zero or negative, the log calculation will fail. Ensure all values remain greater than zero after subtraction.2. Competitive ELISA Constraints: The "Subtract minimum OD value" option is not suitable for competitive ELISAs under a semi-log scale. Because competitive ELISAs feature an inverse relationship between signal and concentration, subtracting minimum OD values easily forces data into negative ranges, crashing the algorithm.
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By default, GainData® calculates final concentrations by first averaging the OD values of your replicates, and then converting that averaged OD into a concentration. This is the standard method to minimize rounding errors.
If you wish to see the calculated concentration for each individual well before averaging, simply assign a different Sample ID number to each well during setup to treat them as independent samples. You can then manually average the resulting concentrations in Excel.
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As a standard laboratory practice, each individual plate must include its own set of standards. Therefore, you should treat them as separate datasets and analyze each plate independently in GainData®.
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The GainData® calculator is designed to compute one standard curve at a time. Since an arigoPLEX® kit detects multiple targets simultaneously (e.g., detecting 4 to 8 cytokines in a single well), you must analyze each target separately. Split your data by target, paste the corresponding high/low standards and sample ODs for that specific analyte, and run separate standard curves sequentially.
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This is usually because your web browser's pop-up blocker is preventing the file from opening or downloading automatically.
【How to Fix】 Check your browser's download folder first. If it isn't there, adjust your browser settings to allow pop-ups for our website. For Google Chrome users, please refer to the Google Chrome Pop-up Support Guide to add our site to the allowed list.
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Currently, GainData® does not support direct export to Word format. To add the standard curve figure to a Word document or lab report, you can take a screenshot (print-screen) of the curve directly from the web results page or extract the image from the generated PDF file.
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This is a temporary glitch caused by local browser caching. Simply press Ctrl + F5 on your keyboard to force-refresh the page and clear the cache. After the page reloads, click "Analyze" and export the data again; the resulting Excel file will open normally.
