protein quantification in cell-based ELISA

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Diana Dragoi
Diana Dragoi's picture
protein quantification in cell-based ELISA

Hello!I am curently performing cell-based ELISA to measure a cell membrane protein in adherently growing cells. I am struggling with the right normalization method. Many cell-based ELISA kits suggest normalization via GAPDH or crystal violet, which do not work for me (treatment of cells influences GAPDH levels; crystal violet produces huge variations/artifacts in samples with few cell numbers).I would prefer to normalize to total protein levels in the same well as I am performing ELISA measurements. Do you have any idea if a BCA-Assay prior to ELISA would interfere with ELISA measurements? I have tried BCA after ELISA (this is currently how it's being done in our lab), but I find this method is not accurate since I am also measuring the amount of antibody bound to the cells.I am also open for alternative methods. Thank you!Diana