ISSN:0275-6382    

DOI:10.1111/j.1939-165X.1991.tb00559.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

ISSN:0275-6382    

DOI:10.1111/j.1939-165X.1991.tb00560.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

SummarySera from two blood type B cats had strong isoagglutinating and isohemolyzing titers against blood type A erythrocytes. In order to determine the class of the immunoglobulins, sera from the cats were pooled, ammonium sulfate precipitated, and gel filtered using sepharose 6B to separate the immunoglobulins by molecular size. The immunoglobulin concentrate separated into two fractions. The initial part of the first fraction was shown in an ELISA to contain IgM and to be devoid of IgG by immunoelectrophoresis and to have agglutinating activity (a titer of 1:4 with a protein concentration of 0.8 mg/ml). Treating this fraction with the reducing agent dithiothreitol (DTT) eliminated agglutinating activity. The latter portion of the second column fraction was shown to contain IgG by immunoelectrophoresis, and to be devoid of IgM by ELISA. Agglutinating activity was also present in the second fraction (a titer of 1:2 with a protein concentration of 1.9 mg/ml); hemagglutinating activity was not decreased by DTT treatment. These studies show that the predominant anti‐A isoagglutinating activity in pooled sera from two blood type B cats is IgM and that some isoagglutinin activity can be associated with Ig

ISSN:0275-6382    

DOI:10.1111/j.1939-165X.1991.tb00561.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

ISSN:0275-6382    

DOI:10.1111/j.1939-165X.1991.tb00562.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

ISSN:0275-6382    

DOI:10.1111/j.1939-165X.1991.tb00563.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

SummaryImmunofluorescence, tube agglutination, and platelet factor‐3 immunoinjury tests for detecting antiplatelet antibody were compared using a heterologous system of equine platelets and rabbit antiequine platelet serum. Platelet immunofluorescence tests were performed using paraformaldehyde‐fixed platelets in suspension as well as in air‐dried smears on glass slides (solid phase). Bright homogeneous, membranous, specific fluorescence was seen in both assays with anti‐immunoglobulin G (IgG) and protein G fluorescein isothiocynate conjugates (FITC‐conjugates). Protein A conjugate gave nonspecific fluorescence irrespective of normal or antiserum treatment. Anti‐IgG and protein G conjugates in suspension immunofluorescence tests with the same antiserum yielded antibody titers of 1:1024 and 1:128, respectively. Similarly, respective titers of 1:512 and 1:64 were obtained with solid phase immunoassay. Platelet suspension assay was slightly better than the solid phase assay. These observations indicated that anti‐IgG was more sensitive than protein G in detecting antiplatelet antibody by fluorescence microscopy, while protein A was ineffective because of its nonspecificity. Chloroquine treatment of platelets failed to reduce the nonspecific fluorescence. Platelet agglutination and platelet factor‐3 tests were relatively less sensitive to detect equine antipl

ISSN:0275-6382    

DOI:10.1111/j.1939-165X.1991.tb00564.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY