We use an instrument combining optical (surface plasmon resonance) and acoustic (Love mode surface acoustic wave device) real-time measurements on a same surface for the identification of water content in collagen and fibrinogen protein layers. After calibration of the surface acoustic wave device sensitivity by copper electrodeposition and surfactant adsorption, the bound mass and its physical properties – density and optical index – are extracted from the complementary measurement techniques and lead to thickness and water ratio values compatible with the observed signal shifts. Such results are especially usefully for protein layers with a high water content as shown here for collagen on an hydrophobic surface. We obtain the following results: collagen layers include 70 ± 20% water and are 16 ± 3 to 19 ± 3 nm thick for bulk concentrations ranging from 30 to 300 μg/ml. Fibrinogen layers include 50 ± 10% water for layer thicknesses in the 6 ± 1.5 to 13 ± 2 nm range when the bulk concentration is in the 46 to 460 μg/ml range.