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In this paper, we examine the relationships between teachers’ subject matter preparation and experience in teaching and their performance on an instrument measuring mathematical knowledge for teaching Algebra 1. We administered the same instrument to two different samples of teachers−high school practicing teachers and community college faculty − who teach the same algebra content in different levels of institutions, and we compared the performance of the two different samples and the relationships between the measured knowledge and their educational and teaching background across the samples. The comparison suggested that the community college faculty possess a higher level of mathematical knowledge for teaching Algebra 1 than high school teachers. The subsequent analyses using the Multiple Indicator Multiple Causes (MIMIC) models based on our hypothesis on the factors contributing to the differences in the knowledge between the two teacher samples suggest that experience teaching advanced algebra courses has positive effects on the mathematical knowledge for teaching Algebra 1 in both groups. Highlighting the positive effect of algebra-based teaching experience on test performance, we discuss the implications of the impact of subject specific experience in teaching on teachers’ mathematical content knowledge for teaching.