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The HMPV monoclonal antibody specifically binds to CD227 which is also known as Mucin-1 (MUC1). A major form of CD227 is expressed as a type I transmembrane glycoprotein. CD227 belongs to the epithelial mucin family whose members are heavily O-glycosylated and characterized by high molecular weight, and an amino acid composition rich in serine, threonine, proline, and glycine. CD227 is variably expressed on the surfaces of normal and malignant glandular and ductal epithelial cells, and some hematopoietic cell lineages including subsets of T cells, B cells, monocytes and dendritic cells. Soluble forms of CD227 may arise by shedding from the cell surface or by secretion of forms derived from alternative RNA splicing. The HMPV antibody binds to the core peptide of the MUC1 protein. The core protein contains a domain of 20 amino-acid tandem repeats which function as multiple epitopes for this monoclonal antibody. Incomplete glycosylation of some tumor-associated mucins may lead to variable unmasking of the multiple peptide epitopes leading to the observed differences in immunostaining intensities between cells from normal and malignant tissues. CD227 plays roles in the provision of protective barrier function, the regulation of cellular adhesion, and the transduction of multiple signal pathways.
FITC, fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.
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