Immunoelectron microscopic detection of band 3 protein during erythroid cell differentiation by a monoclonal antibody.
We created a monoclonal antibody, designated EB1 (IgM, kappa), that reacts with erythroblasts by fusion of P3-X63-Ag8.653 with splenocytes of rats immunized with erythroblastic islands isolated from mice spleens. Western blotting revealed that EB1 reacted with the band 3 protein of the erythrocytic membrane. It stained erythrocytes and erythroblasts, forming clusters in the bone marrow, splenic red pulp, and fetal liver, but did not stain other tissues in the cryostat sections. The EB1 antigen was detected during dimethyl sulfoxide-induced differentiation of murine erythroleukemia cells. Immunoelectron microscopy revealed that the EB1 antigen was expressed from the basophilic erythroblasts during normal erythroid differentiation. Preferential segregation of the EB1 antigen on the cell membrane of the nucleating erythroblasts was not observed. These results suggest that EB1 is specific for erythrocyte band 3 protein and may be useful for studying erythroid cell differentiation.
Sadahira
Y
Y
Department of Experimental Pathology, Kawasaki Medical School, Kurashiki, Japan.
Hirao
Y
Y
Uehira
K
K
Kimoto
T
T
D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000276:immunology* / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D002454:Cell Differentiation; D002460:Cell Line; D015672:Erythroid Precursor Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D004920:Erythropoiesis / Q000276:immunology*; D007150:Immunohistochemistry; D051379:Mice; D016253:Microscopy, Immunoelectron; D051381:Rats; D011919:Rats, Inbred Strains; D013154:Spleen / Q000166:cytology / Q000276:immunology / Q000378:metabolism
D001457:Anion Exchange Protein 1, Erythrocyte; D000911:Antibodies, Monoclonal