PURPOSE:To obtain an analytical element, consisting of a color reaction layer containing a substance capable of reacting plural water-permeable layers with a nondiffusive substance-containing substrate layer having a coloring matter part at a temporarily shortened wavelength and a substrate to provide the coloring matter part at a longer wavelength and useful for immunoassay with high accuracy and sensitivity. CONSTITUTION:A substrate layer 30 containing a substrate which is a nondiffusive substance having a spectral absorption at temporarily shortened wavelength in the molecule and reactive in the presence of an enzymically active substance to produce a diffusive substance having a coloring matter part with spectral absorption at a temporarily shortened wavelength in the molecule and a color reaction layer 20 containing a reactive substance reactive with the diffusive substance produced in the substrate layer 30 to lengthen the wavelength of spectral absorption of the coloring matter part at the temporarily shortened wavelength under the layer 30 as at least two water-permeable layers on a light-transmitting water-impermeable support 10. A contact preventing layer 40, as necessary, is formed between the substrate layer 30 and the color reaction layer 20 or a developing layer 50 is formed on the substrate layer 30 to afford the aimed analytical element useful for immunoassay, etc.