The present study aims to characterize the components and assess the conditions of a deteriorated mural oil painting at Fatma Ismail Palace (Agricultural Museum), Giza, Egypt. Different analytical techniques were used, i.e., the portable digital optical microscope, stereo microscope, Infrared (IR), Ultraviolet (UV), Scanning Electron Microscopy with EDX (SEM.EDX), X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), and Gas Chromatography-Mass Spectrometry (GC MS). The findings illustrated the execution technique and external surface of the mural oil painting. Furthermore, the used microscopes showed various deterioration aspects, including flakes, cleavage, micro-cracks, cracks, as well as detached paint layer and the painting ground from support in some parts. The results of elemental analysis by EDX and XRD showed that the red pigment was red hematite, the yellow pigment was yellow ochre, the blue pigment was ultramarine blue (Lazurite), the green pigment was earth green (Glauconite), and the black pigment was carbon black (Graphite). The analysis of soluble salts showed that NaCl was the most common soluble salt. FTIR analysis determined the medium employed as a binder in the painting ground and plaster layer. It showed the functional group of the plaster layer containing gypsum CaSO4. H2O without a binder and that of the painting ground consisting of Calcite (CaCO3), barite (BaSO4), glue, and oil. GC/MS analysis revealed that walnut oil was used as a medium for the pigments. The present paper concluded significant findings about the analytical methods of assessing the conditions and identifying the components of the mural oil painting. It showed that the mural oil painting understudy suffered from several deterioration aspects. It recommends that further studies should tackle the conservation steps, i.e., conservation treatment or preventive conservation.