Organic shale beds in the Alam El-Bueib (AEB) are thought to provide an excellent source for producing hydrocarbons. Consequently, defining the role of the AEB formation in hydrocarbons generation in the Tut Oil Field is the primary objective of this research. This research presents an analysis of biomarkers in a set of Tut Oil Field AEB extracts and oils to appreciate the basin's hydrocarbons xploration and development. AEB shales have low Pr/Ph ratios (0.5e0.83), low to high values of C22/C21 tricyclic terpanes, low values of C19/C23 tricyclic terpanes, high values of C31 R homohopanes/C30 hopanes, and high concentration of regular steranes C27 decode that the organic matter was primarily derived from marine algae/bacterial derived organic matter, along with small amounts of land plants formed in a reducing environment. In addition, the maturity-related parameters display values ranging from low to high. The oils recovered from Bahariya and AEB reservoirs have higher Pr/Ph ratios, and high values of tricyclic terpanes (C22/C21) decode that the organic matter was primarily derived from marine algae/bacterial-derived organic matter, and an elevated concentration of C29 steranes interpret that the organic matter was primarily derived from mixed marine algae/land plant-derived organic matter formed in a suboxic environment. The maturity-related parameters display high maturity values. Based on various biomarker criteria, the hierarchical cluster analysis of crude oils and AEB extract reveals no genetic relationship between the AEB source rock and the studied oils.