The study was conducted to investigated effect of three amino-acids which were Methionine(0.2 Mm), L-Arginine (0.4 Mm) and D-Alanine(15 Mm) on Awassi epididymal sperms characteristics after storage at 4Co. Twenty five pairs of testicles (n=25, 50 testis/semen samples ) from 25 mature Awassi Rams aged between 1-4 years were used in present study. Epididymal sperms were collecting by slicing of epididymis and diluted in egg yolk extender.Four test tubes samples were created, each of one contained an equal amount of antibiotic, fructose, one kind of amino acid, sodium citrate 2.9%, and egg yolk10%. The fourth test tube left as control group and all test tubes keeps in refrigerator temperature at 4Co. The results show that after 24,27 hours of storage, Methionine improve significantly p<0.05 sperm individual motility which were 72.6 ± 2.9, 57.5 ± 3.2 respectively, sperm live percentage was 73.6 ± 2.3, 53.2 ± 2.8 respectively, Sperm abnormalities was 3.0 ± 0.1, 7.2 ± 0.1 respectively. Arginine was following Methionine effect on sperm individual motility which were 66.0 ± 3.3, 47.0 ± 3.8 respectively, live percentage was 70.1 ± 2.8, 49.2 ± 3.3 respectively, Sperm abnormalities was 4.0 ± 0.1, 7.7 ± 0.1 respectively. Alanine data were significantly less effect on sperm individual motility which were 64.0 ± 3.1, 45.6 ± 3.2 respectively, sperm live percentage was 65.8 ± 3.0, 46.8 ± 3.2 respectively, in conclusion, Arginine (0.4 Mm) and Alanine (15 Mm) also act to improve sperm characteristics when compare with control group in Awassi rams epididymal sperms.