Gly m Bd 30K protein is recognized as a major immunodominant allergen in soybean， and this protein can lead to anaphylaxis such as diarrhea and intestinal inflammation in humans and livestock. The exploration of germplasms with lower Gly m Bd 30K content is important in breeding for elite functional soybean varieties. In this study， we generated the polyclonal antibody targeting to the 190-379aa peptides of the Gly m Bd 30K protein， followed by quantifying the Gly m Bd 30K protein using Western blot in 30 soybean accessions that were derived from Shanxi province. The genomic sequence of the Gly m Bd 30K gene and its transcriptional level was further conducted. As a result， two germplasms（ 134， ZDD02046， Daqingdou， ZDD02174） with lower Gly m Bd 30K protein content have been identified， the identification efficiency is 6.9%. Both accessions displayed 8 and 42 TA repeats respectively in the promoter of the Gly m Bd 30K gene if compared with that of Willams82. The transcription level in 134（ ZDD02046） was found to be significantly lower than that of Daqingdou（ ZDD02174） . It is speculated that TA polymorphism in the promoter may associate with the Gly m Bd 30K expression. Collectively， this study established a strategy on the quantification of Gly m Bd 30K protein content and identified two accessions with lower Gly m Bd 30K protein， which will provide technical and material supports in breeding for new soybean varieties with high-quality proteins.