Abstract: The ‘Hariana’ breed of cattle has been proved to be highly adaptable to harsh climatic conditions and resistant to common diseases prevalent in India. In this study, the normal reference values of hematological and serum proteins and other blood biochemical parameters were determined in the heifers of Hariana breed maintained at Instructional livestock farm complex, DUVASU, Mathura, India. A total of twenty four animals were used in this study. Blood was taken aseptically from all the animals and transported to laboratory for hematological and biochemical analysis. The hematological parameters (Hemoglobin, total erythrocyte count, total leukocyte count, packed cell volume) and biochemical parameters (Total protein, total albumin, albumin globulin ratio, urea, creatinine, calcium, phosphorous, calcium phosphorous ratio, AST, ALT) values were statistically analyzed, mean and standard deviations were calculated and set as reference values. This study reported hematological and serum biochemical values which could serve as baseline information for comparison in conditions of nutrient deficiency, physiological and health status of Hariana cattle heifers in India.