Role of antisperm antibodies in infertility of animals is still controversial. A study was planned with an objective to detect antisperm antibodies (ASA) by Immunoperoxidase assay (IPA), Sperm Mar test and ELISA in blood serum and cervical mucus of 53 cross-bred cows, which were interrelated with age, parity and number of inseminations. Cows were grouped according to age (years), parity- and insemination- number as G-1(< 3, 0 and 0), G-II (3.0 - 4.5, 1.0 and 1- 3), G-III (4.6 - 6.0, 2.0 and 4.0 – 6.0) and G-IV (> 6.0, ≥ 3.0 and > 6.0). In Sperm Mar test, 40% reaction between motile spermatozoa and coated latex particles of IgG and IgA is considered as lower limit of significant activity. Therefore, percentage of cows with higher level of ASA (> 40% IPA / IgG / IgA, 3200 - 6400 / 200 - 800 titre) was calculated in each group. Proportion of cows with higher serum-IgG and ELISA-titre in cervical mucus and that with higher ELISA titre in serum / cervical mucus increased with increased age and parity, respectively. According to number of inseminations, percentage of cows with higher  level of ASA (IPA), IgG and ELISA titre in blood serum was maximum in G-IV, whereas that with IgA in G-III  in comparison to other groups. Percentage of cows with higher level of ASA (IPA), IgG, IgA and ELISA in cervical mucus was higher in G-III as compared to other groups. This study exposed a significant increase in ASA in serum /cervical mucus of cross-bred cows only with increase in number of inseminations.