Abstract: Anti-globulin reagent is the most important tool used in immunohematology laboratories and blood banks for allo-antibodies screening during incompatible pregnancy and cross matching. this study, attempts were made to prepare a suitables monospecific reagent, for Rh-D allo-immunization during pregnancy; by immunizing rubbits with fresh human serum and purified human immunoglobulin (Ig) anti-human immunoglobulin and anti-human serum were produced and used at a dilutions 1/120 and 1/100 , respectively. No significant differences were found in using both the prepared reagent, for antibody screening during pregnancy from the commercial anti-globulin reagent . Local polspecific anti-globulin reagent was prepared by mixing the prepared anti-complement with the prepared anti-human (Ig) in appropriate dilution for detection of human immunoglobulin and complement coated red blood cells. The locally prepared polyspecific anti-Ig globulin reagent was used for antibody screening during cross matching in a comparison with the prepared anti-human serum and the commercial anti-globulin reagent. The prepared anti-human show significant difference from the commercial AHG reagent in cross matching, while no significant difference was observed when the locally prepared polyspecific anti-globulin reagent was used in comparison with the commercial reagent in cross matching . New serological test for antibody screening, represented by solid phase anti-globulin test (SPT) , have been described, and the SPT level of sensitivity was comparable to that of the manual indirect anti-globulin test (IAGT) for the detection of IgG antibodies. The data demonstrated a linear and overlapping relationship between the score of manual IAGT and SPT optical density values . The optical density value of SPT increased as the concentration of RBC increased. The optimum concentration of immunoglobulin used in the preparation of- solid phase was- about 1 mg/ml . The cutoff point of negativity was determined (0.021 ± 0.0107) when samples from healthy Donors were tested.