ELISA & Protein Assay Analysis
Paste the plate readings, mark blanks, standards and samples on the plate map, and get a fitted standard curve with sample concentrations — corrected for dilution and flagged when they fall outside the standards. Protein assay mode adds a calculator for equal loading on a Western blot.
Runs in your browser · readings are never uploaded · 96-well platesLoading… (needs JavaScript)
How to use it
1. Choose ELISA or Protein assay. 2. Paste the readings — the 8 × 12 block from the plate reader export or a spreadsheet; optionally paste the reference-wavelength block too. 3. Check the plate map: B for blanks, S1, S2 … for standards, U1, U2 … (or any name) for samples; wells with the same code are replicates. 4. Enter standard concentrations (or fill a serial dilution) and each sample’s dilution factor. 5. Pick the curve model and read the results, standard recovery and flags; download the CSV, the curve and a methods paragraph.
What it computes
Reference readings are subtracted well by well and the mean blank is subtracted from every well. Standards are fitted by least squares: the four- or five-parameter logistic (Levenberg–Marquardt), a straight line, a second-order polynomial or a log–log line, optionally weighted by 1/y². Each sample’s mean reading is converted to concentration with the inverse of the fitted curve and multiplied by its dilution factor. For every standard the tool reports the back-calculated concentration and recovery (80–120 % is the usual acceptance range) and the replicate CV. Samples below the lowest non-zero standard or above the highest are flagged rather than reported as quantified. In protein assay mode the loading calculator gives the lysate, sample-buffer and water volumes for the same protein mass in every lane.
Limits
Report only concentrations inside the standard range; re-run out-of-range samples at another dilution. The 5PL needs standards that cover both plateaus — if its parameters run away the tool warns you, and the 4PL is the safer choice. Standards and samples should be in the same matrix and buffer, and colorimetric protein assays are affected by detergents, reducing agents and chelators in the lysis buffer — check your reagent’s compatibility table. Replicate wells of one sample are technical replicates.
Keep your work
Nothing is uploaded or stored. Download the results CSV and the settings file (which holds the readings and plate map) before closing the page.
Privacy and local-data controls · Credits and licenses · Interpretation limits