Boosting underrepresented student persistence in STEM majors has been of interest for several years. Prior research has explored various factors that may influence student success and persistence in STEM majors. Specifically, some of these studies have employed Yosso's community cultural wealth framework to explore how a student's cultural wealth may link to certain outcomes. Most of these studies have taken a qualitative approach when exploring cultural wealth and less adopt a quantitative approach. Using biology students in an introductory biology class during the fall (n = 303) and spring semesters (n = 215) at a large Hispanic-serving institution in the southeastern United States, this study seeks to contribute to the literature by validating a previously constructed instrument with a new population. An exploratory factor analysis (EFA) was conducted using principal axis factoring and an oblique rotation. EFA results revealed 10 dimensions of cultural wealth. Additionally, confirmatory factor analysis confirmed that the model produced properly measures the constructs as intended. Overall, the final 56-item instrument used in this study was validated and can be used for measuring cultural wealth in similar populations.