The Chen’er gold deposit, with 27 tons of known Au reserves, is one of the typical gold deposits in the Xiaoqinling district along the southern part of the North China Craton. In order to better understand the gold precipitation time and mineralization process, we report in-situ Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) trace element analyses, elemental maps, and the Rb-Sr and S-Pb isotopic composition of sulfides from the Chen’er gold deposit with a view to gain insights into the mineralization time and process. Trace element analyses and elemental maps of pyrite show that the content of As is low and has no correlation with Au, indicating As is unlikely to have controlled the Au incorporation into pyrite. Gold mainly occurs as solid solutions in the Py Ⅰ and Py Ⅱ, and as Au-Ag and/or Au-Te sub-micrometer inclusions in Py III. The Rb-Sr isotopic data on sulfides from the Chen’er deposit yielded an isochron age of 129.2 ± 3.4 Ma, which overlaps with the previously reported 40Ar/39Ar age of hydrothermal mica, indicating that the gold mineralization occurred in the Early Cretaceous. The in-situ sulfur isotope values of sulfides from stage Ⅱ to stage III fluctuate significantly (the average value decreases from 4.95 to −0.24‰) and were affected by physicochemical parameters of the mineralization process, especially ƒO2. In addition, the δ34S values are in a narrow range (-1.34 to 5.24‰), indicating a magmatic source for the sulfur. In-situ Pb isotopic compositions of the pyrite and galena of stage Ⅱ and stage III are relatively constant with 208Pb/204Pb, 207Pb/204Pb, and 206Pb/204Pb values in the range of 37.264–37.595, 15.364–15.423, 16.992–17.078, respectively, indicating a mixed mantle-crust origin. Based on the new geochronological, geochemical and isotopic data presented in this study, and combined with those from previous studies, we suggest that the lode gold deposits in the Xiaoqinling district are magmatic hydrothermal ore deposits, with metal and ore-forming fluid originating primarily from the mantle, coupled with the cratonic reactivation during the Early Cretaceous.