Qualitative research conducted through occupational socialization theory has documented that physical educators feel marginalized and isolated in schools. We sought to propose and provide initial evidence of validity and reliability for the Physical Education Marginalization and Isolation Survey (PE-MAIS). Physical educators (n = 420) completed an online survey including isolation and marginalization, perceived mattering, and burnout. Data analyses began with exploratory factor analysis, which identified a stable 10-item, 2-factor structure. Confirmatory factor analysis was used to affirm the structure and examine convergent, discriminant, and divergent validity. The model fit was good, X[superscript 2](34) = 71.12, p < 0.001, NNFI = 0.91, CFI = 0.94, SRMR = 0.05, RMSEA = 0.07 (90% CI [0.052, 0.093], p = 0.03), and the new measure correlated positively with burnout and negatively with perceived mattering. Invariance analysis indicated that elementary and secondary physical educators interpreted the constructs similarly, and there were no significant differences between the groups.