We have identified a small proline-rich protein, spr1, which is a sensitive and specific marker for distinguishing squamous cancer from the other cell types of lung carcinoma. A rabbit antiserum against a 15-amino-acid peptide of the C-terminus of spr1 was prepared. The specificity of this antiserum was demonstrated in normal squamous tissues by Western blotting and immunohistochemical analysis. Expression of spr1 in 63 cases of formalin-fixed and paraffin-embedded human bronchogenic carcinoma was studied by immunohistochemical staining. For these 63 cases of bronchogenic carcinoma previously diagnosed by hematoxylin/eosin (H/E) staining, the number of spr1-positive cases/total number of H/E cases of each cell type of lung cancer were as follows: 20/20 of squamous carcinoma, 2/18 of adenocarcinoma, 4/14 of large-cell carcinoma, and 0/11 of small-cell lung carcinoma. Squamous differentiation evidenced by spr1 expression was substantiated by the presence of squamous features observed under transmission electron microscopy (TEM). We conclude that spr1 is a sensitive and specific marker for squamous bronchogenic carcinoma.