Background and Objectives Red cell antibody screening as a part of pretransfusion testing is not a routine practice in many developing countries. This study, was done to determine the incidence of allo-and auto-antibodies, assess their clinical significance and deliver antigen negative blood where warranted.Materials and Methods Red cell antibody screening was done as a part of pretransfusion testing at a tertiary care multispecialty hospital in India on 60 518 patients between August 2007 and July 2014 using column agglutination technology by gel.Results Red cell antibodies were detected in 0Á95% (n = 579). Of these, 0Á54% (n = 332) were alloantibodies and 0Á4% (n = 247) were auto-antibodies. Rh and Kell antibodies detected were anti-D(n = 140), anti-C(n = 10), anti-c(n = 12), anti-E(n = 21), anti-e(n = 4), anti-K(n = 10), anti-k(n = 2). Anti-M (n = 30), anti-N(n = 1), anti-S(n = 7), anti-Le(a)( n = 23), anti-Le(b)( n = 6) and anti-Fy(a) (n = 7) were found. Dual antibodies anti-D&C(n = 36), anti-C&E(n = 1), anti-c&E (n = 2), anti-K&C(n = 1), anti-Jka&E(n = 1) and anti-Jka&D(n = 1) were also detected. Of the 332 alloantibodies detected only 9 could not be typed. Additionally, our study detected 247 auto-antibodies.Conclusion Implementation of red cell antibody screening routinely helped us understand the prevalence of antibodies in our region and showed that wider implementation at the national level is feasible and adds to blood safety.