The study was undertaken to investigate biochemical diversity within and between Tiv and Fulani local chicken ecotypes of Nigeria. The experimental birds were randomly sourced from ten locations. The locations (1-5) for the Tiv ecotype were Uikpan, Daudu, Kadarko, Yelwata and Cohor (in Benue and Nasarawa States) while that of the Fulani ecotype were Lafia, Akurba, Adogi, Asakio and Namu (in Nasarawa and Plateau States). At maturity, four (4) male and four (4) female birds were randomly selected from each location per ecotype to give a total of eighty (80) adult birds (40 birds each for Tiv and Fulani ecotype) and used for blood protein characterization study. Blood protein loci, namely haemoglobin, albumen, transferrin and carbonic anhydrase were analyzed using electrophoresis. Data collected from the biochemical analysis were analyzed using popgene version 1.31. The results indicate that, Allele A was prevalence in both the Tiv (0.563) and the Fulani (0.769) ecotypes for heamoglobin while Allele C for albumen was most prevalent in both ecotypes. In transferrin however, allele D (0.419) and allele A (0.689) were most prevalent in the Tiv and the Fulani ecotype, respectively. Carbonic anhydrase showed that Allele A dominated in both ecotypes with frequencies of 0.550 and 0.632 for the Tiv and the Fulani ecotypes respectively. The genetic variability (heterezygosity) value of the two ecotypes ranged from 0.360 to 0.654. The Tiv ecotype had higher heterozygosity value at all loci than the Fulani ecotype. All the protein loci for the two ecotypes had 100 percent polymorphic loci except in the Fulani ecotype where 75 percent polymorphic loci were observed for locations 2 and 3. The effective number of alleles (ne) was 2.273 and 1.779 for the Tiv and the Fulani ecotype, respectively with an average of 2.026. From the findings of this study, it was concluded that the Tiv and the Fulani chicken ecotypes are distinct genetic groups with sufficient genetic variability within