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  • Preparation of a monoclonal antibody to citrullinated epitopes: its characterization and some applications to immunohistochemistry in human brain. 11870872

    Using hybridoma technology, an IgM monoclonal antibody (mAb), designated as F95, was developed against a deca-citrullinated peptide (DCP) consisting of 10 citrulline residues and a carboxyl Gly-Gly-Cys through which DCP was covalently linked to an activated carrier protein, keyhole limpet hemocyanin (KLH). Clones were selected on the basis of not reacting with human unmodified and noncitrullinated myelin basic protein (MBP), MBP-C1, but reacting well with human citrullinated MBP (MBP-C8). When tested by ELISA, this mAb demonstrated minimal reactivity with human MBP-C1, varying reactivity with the C2-C5 isomers of human MBP, moderate binding with guinea pig MBP-C8, and strong reactivity with human MBP-C8. By ELISA, mAb F95 was directed predominantly against citrulline, not MBP, as revealed by its binding to DCP linked with activated KLH, bovine serum albumin (BSA), or ovalbumin (OA), but not with KLH, BSA, or OA alone. Immunohistochemistry of normal human brain demonstrated that F95 stained central nervous system myelin and a subset of astrocytes. Given the citrulline-directed features of mAb F95, this immunohistochemical pattern suggests that certain astroglial filaments expressing glial fibrillary acidic protein also contain citrulline-bearing components. These potentially implicate citrullinated proteins, notably in astroglial filaments, in a variety of normal and pathological neurobiological processes.
    Tipo de documento:
    Referencia
    Referencia del producto:
    MABN328
    Nombre del producto:
    Anti-peptidyl-citrulline, clone F95 Antibody

  • Molecular characterization of the dimer formation of Fcα/μ receptor (CD351). 23665380

    Fcα/μR (CD351) is an Fc receptor for both IgA and IgM and forms an atypical dimer that is resistant to reduction by 2-mercaptoethanol or boiling. We previously demonstrated that the cytoplasmic portion of Fcα/μR is required for dimer formation and for its efficient cell-surface expression. However, the biochemical nature of these phenomena has not been determined. By using a BW5147 mouse cell line expressing deletion mutants of the cytoplasmic region of Fcα/μR, we found that the region spanning amino acids 504-523 was required for efficient cell-surface expression, whereas the region spanning amino acids 481-490 was required for dimmer formation. Immunoblotting analyses of transfectants simultaneously expressing Flag-tagged Fcα/μR and hemagglutinin-tagged Fcα/μR suggested that Fcα/μR does not form homodimers. Instead, our data suggest that Fcα/μR forms heterodimers with an as-yet-unknown molecule with a molecular weight of 60-70 kDa.
    Tipo de documento:
    Referencia
    Referencia del producto:
    Múltiplo
    Nombre del producto:
    Múltiplo

  • Human remyelination promoting antibody inhibits apoptotic signaling and differentiation through Lyn kinase in primary rat oligodendrocytes. 20645409

    Human remyelination promoting IgM mAbs target oligodendrocytes (OLs) and function in animal models of multiple sclerosis (MS). However, their mechanism of action is unknown. This study seeks to identify the cellular mechanism of action of a recombinant human IgM on OL survival.Binding of rHIgM22 to the surface of rat OLs was studied by co-localization with various markers. RHIgM22-mediated effects on apoptotic signaling in OLs, differentiation markers, and signaling molecules were detected by Western blotting and immunoprecipitation.RHIgM22 co-localized with integrin β3 but not other integrin β-chains in OLs. Downstream of integrin β3 we identified Src family kinase (SFK) Lyn as a key player of rHIgM22-mediated actions in OLs. Lyn immunoprecipitated in a complex together with integrin αvβ3 and PDGFαR. Lyn expression was 9-fold up-regulated and Lyn activation was 3-fold higher inrHIgM22-treated OL cultures compared with controls. RHIgM22 inhibited apoptotic signaling by greater than 10-fold reduction of caspase-3 and capsase-9 cleavage and reduced by 4-fold expression of differentiation markers MBP and MOG in OLs. SFK inhibitors PP2 and SU6656 inhibited Lyn activity and restored caspase-cleavage in OLs. A human IgM that did not promote remyelination and medium wereused as controls.rHIgM22 prevented apoptotic signaling andinhibited OL differentiation by Lyn implying thatIgM-mediated remyelination is due toprotection of OPC and OLs rather than promotion of OPC differentiation.
    Tipo de documento:
    Referencia
    Referencia del producto:
    Múltiplo
    Nombre del producto:
    Múltiplo

  • Comparative studies on monoclonal antibodies raised against human gastric cancer for application to serum diagnosis of cancer. 2455469

    Four murine MoAbs, KM191(IgM), KM206(IgM), KM230(IgG1) and KM231(IgG1), against human gastric cancer were generated using mice which underwent tolerance treatment to stomach tissues. They exhibited very similar high reactivities to stomach adenocarcinoma cells and low reactivities to normal cells in both membrane binding assay and immunohistochemical analysis. Their antigens were neuraminidase and protease sensitive and existed as macromolecules (1,000Kd) in body fluids. Binding of each antibody was inhibited by the others and KM231 showed the highest binding avidity. When they were used to detect the antigens shed in ascitic fluids and pleural effusions of cancer patients, KM231 allowed the most efficient detection of the antigen. The above results indicated that the four MoAbs bound to closely related epitopes on the same antigen and that the nature of its high binding avidity enabled KM231 to show the greatest efficiency in the detection of the antigen in body fluids. KM231 was applied to serum diagnosis and gave high positive percentages in pancreatic cancer(86%), hepatocarcinoma(87%), gall bladder cancer(50%), and gastric cancer(34%), whereas in healthy persons (0%) and benign diseases except for hepatitis(29%) the percentages were low. KM231 was similar to NS19-9, but quite different from NS19-9 in the high positive percentages of hepatocarcinoma in serum diagnosis.
    Tipo de documento:
    Referencia
    Referencia del producto:
    MAB2095