Immunoblot Analysis of CD34 Expression in Histologically Diverse Neoplasms

Stanford University, Palo Alto, California, United States
American Journal Of Pathology (Impact Factor: 4.59). 02/2000; 156(1):21-7. DOI: 10.1016/S0002-9440(10)64701-3
Source: PubMed


CD34 is a heavily glycosylated transmembrane protein of approximately 110 kd whose function is essentially uncharacterized. First identified in a myeloid leukemia cell line, immunohistological reactivity with anti-CD34 antibodies is also encountered in a histologically diverse subset of nonhematolymphoid neoplasms including angiosarcoma, solitary fibrous tumors, epithelioid sarcomas, spindle cell lipomas, dermatofibrosarcoma protuberans, and myofibroblastomas. Immunohistological reactivity for CD34 in hematopoietic stem cells and endothelial cells has been shown to correspond to the expression of the CD34 protein. With the exception of gastrointestinal stromal tumors, CD34 protein expression has not been investigated in other CD34 immunohistologically reactive nonhematolymphoid neoplasms. We undertook this study to examine whether the observed reactivity for anti-CD34 antibodies in apparently unrelated tumors is due to the expression of the same protein or whether shared epitopes elaborated by other proteins could account for this reactivity. Immunoblot analyses with anti-CD34 antibodies of six different CD34 immunohistologically reactive lesions show the same approximately 110-kd molecular weight protein. In addition, two cases of dermatofibrosarcoma protuberans show double bands at approximately 110 kd. Laser-capture microdissection of CD34 immunohistologically reactive epithelioid sarcoma and nonreactive epidermal cells illustrates that this reactivity is specific to tumor cells. These results show that the observed immunohistological reactivity with anti-CD34 antibodies is due to the expression of the CD34 protein and not to shared epitopes on unrelated proteins.

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    • "However, SDS is not compatible with downstream analytical techniques such as mass spectrometry, unless preceded by gel separation. Gel electrophoresis: Polyacrylamide gel electrophoresis (PAGE) can separate proteins according to their molecular weight and can be followed by Western Blot analysis for highly specific protein characterization [57] [58]. However, this approach is limited by the availability of appropriate antibodies and by the amount of proteins necessary to obtain a signal . "
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