Before investigating any potential between-group differences, it is necessary to test the measurement invariance between groups. Some researchers discussed the measurement invariance of first-order factor model, but the measurement invariance research of second-order factor model was rare. Compared with first-order factor model, the second-order factor model has several advantages, but testing measurement invariance of second-order factor model was more complex. In the paper, the measurement invariance across regions in the Internet Altruistic Behavior Scale of Undergraduates (IABSU) was tested. IABSU which include 26 items comprised four first-order factors and single second-order factor. Items were rated on a 4-point Likert-type scale ranging from 1 (“never do”) to 4 (often do). A total of 592 college students from five universities in Shanghai City and Anhui Province were examined using random sampling method. The 283 college students from Shanghai were taken as the developed region sample, the mean age was 20.1 (SD=1.2). And the 309 college students from Anhui Province were taken as the developing region sample, the mean age was 20.4 (SD=1.1). First, baseline model was established to fit the data of the developed region and the developing region separately. The results showed that the absolute index RMSEA and SRMR were less than .08, the relative index NNFI and CFI were more than .95, which indicated that the model could extremely fit the data. And then, the measurement invariance of second-order factor model of IABSU was tested, following the general procedures suggested by Widaman and Reise (1997), including configural invariance, first-order weak invariance, second- order weak invariance, first-order strong invariance, second-order strong invariance, second-order strict invariance and first-order strict invariance. After the lower level invariance was achieved, the higher level invariance which was more stringent could be tested. All procedures were based on the analysis of mean and covariance structures (MACS) within the framework of CFA modeling. The results showed that IABSU was full first-order and second-order strict invariance across regions. In the paper, we showed clearly how to test the measurement invariance of second-order factor model taking IABSU as an example, and also indicated the measurement invariance across regions of IABSU was good, which could be used to test the internet altruistic behavior in different regions.