Abstract Background Social support represents the belief that one is valued, cared for, and loved by others in a social network and a lack of it may lead to adverse outcomes such as a relapse into depression or emotional distress in physically ill patients. The Multidimensional Scale of Perceived Social Support (MSPSS) is an instrument aimed to measure an individual’s perception of how much he or she receives outside social support. This study aimed to examine the validity and reliability of the Serbian version of MSPSS in a sample of LGBT population in Serbia. Methods The cross-sectional study included 504 respondents, members of the LGBT population living in Serbia. The instrument used was the MSPSS as a self-report measure, containing twelve items rated on a 7-point Likert scale. MSPSS consists of three sub-scales: Family, Friends, and Significant Others. Exploratory (EFA) and confirmatory factor analyses (CFA) were performed with two independent random samples. Results The average score on the scale of social support was 5.58±0.84 (scale of social support by family 4.62±1.41, scale of social support by friends 5.93± 1.99, and scale of social support by the significant others 6.20±0.96, respectively). The Cronbach’s alpha for the entire social support scale was α = 0.772. The three-factor structure of the MSPSS was proved with EFA and CFA. The factor loadings varied between 0.630-0.817. The three factors explained a total of 52.68% of the variance (factor 1- 28.62%, factor 2. 13.16%, factor 3- 10,90%). The three-factor model was adopted during the CFA when the three-factor model CFI (comparative fit index) was 0.969. Conclusions The MSPSS has shown good reliability, internal validity and construct validity and can be used as a convenient measure of social support among the LGBT population. Both EFA and CFA yield a three-factor structure consistent with the theory underlying the measure’s development. Key messages • The results of the study suggest that the Multidimensional Scale of Perceived Social Support has acceptable psychometric properties for use with the LGBT population. • This study supports the use of the MSPSS for assessing community-based collaboration using the three-factor model among the LGBT population.