Calpastatin (CAST) is a specific inhibitor of calpain (calcium-dependent cysteine protease). This study investigated the potential for variation in yak (Bos grennies) CAST. PCR-SSCP analysis of exon 6 of yak CAST revealed three unique patterns (named A-C). Sequencing of the amplicons revealed two nucleotide substitutions. One substitution (c.398G/C) would nominally change the amino acid sequence (p.S133T) of yak calpastatin. The variant sequence A which carried c.398C was the most common in the yaks tested (95.1%). This is the first report that found yak CAST is variable, and as in pigs, sheep and cattle, this variation may affect animal production traits.