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Protein content is one of the main factors determining the end-use quality of wheat. <i>NO APICAL MERISTEM-B1</i> (<i>NAM-B1</i>) is a major gene regulating wheat grain protein content. The present study aimed to identify new genetic resources using the wild-type <i>NAM-B1</i> allele to breed high-protein-content wheat cultivars. We genotyped the <i>HIGH GRAIN PROTEIN CONTENT-B1</i> (<i>GPC-B1</i>) locus and <i>NAM-B1</i> allele in 165 wheat cultivars. A kompetitive allele-specific polymerase chain reaction (KASP) marker was designed for functional <i>NAM-B1</i> allele screening. The results revealed that 41 out of 165 cultivars carried the <i>GPC-B1</i> locus. Among the 41 <i>GPC-B1</i>-carrying cultivars, the wild-type <i>NAM-B1</i> allele was identified in only 3 cultivars, none of which were Korean. The remaining 38 cultivars showed a 1-bp insertion in <i>NAM-B1</i>, resulting in a stop codon in the middle of the gene, rendering it nonfunctional. Overall, this study reveals that the utilization of the three selected cultivars possessing the wild-type <i>NAM-B1</i> gene, in conjunction with the developed KASP assay, could increase the protein content in Korean wheat cultivars.