Semi-Automated Rapid Isoelectric Focusing of Apolipoproteins C from Human Plasma Using PhastSystem™ and Immunofixation

Noll, Bernd; Hackler, Rolf; Pelzer, Mirko; Pelzer, Sabine; Nusser, Petra; Maisch, Bernhard; Schaefer, Juergen R.; Steinmetz, Armin

doi:10.1515/cclm.1999.100

Cited by 9 publications

(5 citation statements)

References 28 publications

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“…Analyses of serum Tf and aAT [33], ApoE [34] and ApoCIII [35] were performed by isoelectric focusing (IEF) with direct immunofixation and Coomassie or silver staining. For proper identification of protein variants, digestion with neuraminidase (from Clostridium perfringens, Sigma, St. Louis, USA) prior to IEF was used [33].…”

Section: Methodsmentioning

confidence: 99%

Microheterogeneity of some serum glycoproteins in neurodegenerative diseases

Marklová

Albahri

Vališ

2012

Journal of the Neurological Sciences

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Section: Methodsmentioning

confidence: 99%

Microheterogeneity of some serum glycoproteins in neurodegenerative diseases

Marklová

Albahri

Vališ

2012

Journal of the Neurological Sciences

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“…Automated IEF of serum samples and detection of a 1 -antitrypsin and a 1 -antichymotrypsin was performed according to Hackler et al (in preparation) using a Phast-System TM . Neuraminidase treatment of serum was performed according to Noll et al [32].…”

Section: Methodsmentioning

confidence: 99%

Congenital Disorder of Glycosylation IId (CDG-IId) - A New Entity: Clinical Presentation with Dandy-Walker Malformation and Myopathy

Peters¹,

Penzien²,

Reiter³

et al. 2002

Neuropediatrics

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A 1.5-year-old boy with macrocephaly due to a Dandy-Walker malformation presented with progressive hydrocephalus, extensive muscular hypotonia, transient cholestatic syndrome, extensive coagulation abnormalities and elevated creatine kinase indicating myopathy. Diagnostic work-up indicated a congenital disorder of glycosylation (CDG, formerly carbohydrate deficient glycoprotein syndrome). The serum transferrin pattern obtained by automated isoelectric focusing (IEF) showed an hitherto unreported pattern with strongly elevated tri-, di-, mono- and asialotransferrin bands, increasing in this order together with markedly decreased tetrasialotransferrin. Investigation of two additional glycoproteins, alpha(1)-antitrypsin and alpha(1)-antichymotrypsin, confirmed a generalised defect of glycosylation. All known glycosylation defects could be ruled out by enzymatic analyses in either leukocytes or fibroblasts or by the results obtained by IEF. SDS-electrophoresis demonstrated a marked difference in the molecular weight of transferrin, suggesting the lack of parts or of all oligosaccharide chains. The defect could be delineated to a deficiency of beta-1,4-galactosyltransferase (E.C.2.4.1.38) due to a homozygous insertion (1031 - 1032 insC). Details of the biochemical and molecular findings will be described elsewhere.

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“…After 6 h, serum from BALB/c mothers, fetal serum, and amniotic fluid were collected. Isoelectric focusing of serum and amniotic fluid was performed in polyacrylamide gels (pH 4.0 -6.5) using the PhastSystem (Amersham Pharmacia Biotech, Freiburg, Germany), followed by immunofixation as described previously (16). Immunofixation was performed by incubation with either polyclonal antimouse IgG serum (anti-OVA serum) or nonimmune mouse serum (control serum).…”

Section: Determination Of Allergen Transfer To the Fetusmentioning

confidence: 99%

Critical Role of Preconceptional Immunization for Protective and Nonpathological Specific Immunity in Murine Neonates

Uthoff

Spenner

Reckelkamm

et al. 2003

The Journal of Immunology

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111

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Expression of Th2 immunity against environmental Ags is the hallmark of the allergic phenotype and contrasts with the Th1-like pattern, which is stably expressed in healthy adults throughout life. Epidemiological studies indicate that the prenatal environment plays an important and decisive role in the development of allergy later in life. Since the underlying mechanisms were unclear, an animal model was developed to study the impact of maternal allergy on the development of an allergic immune response in early life. An allergic Th2 response was induced in pregnant mice by sensitization and aerosol allergen exposure. Both, IgG1 and IgG2a, but not IgE, Abs cross the placental barrier. Free allergen also crosses the placental area and was detected in serum and amniotic fluids of neonatal F1 mice. These F1 mice demonstrated a suppressed Th1 response, as reflected by lowered frequencies and reduced levels of IFN-γ production. Development of an IgE response against the same allergen was completely prevented early in life. This effect was mediated by diaplacental transfer of allergen-specific IgG1 Abs. In contrast, allergic sensitization against a different allergen early in life was accelerated in these mice. This effect was mediated by maternal CD4 and OVA-specific Th2 cells induced by allergic sensitization during pregnancy. These data indicate a critical role for maternal T and B cell response in shaping pre- and postnatal maturation of specific immunity to allergens.

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Semi-Automated Rapid Isoelectric Focusing of Apolipoproteins C from Human Plasma Using PhastSystem™ and Immunofixation

Cited by 9 publications

References 28 publications

Microheterogeneity of some serum glycoproteins in neurodegenerative diseases

Microheterogeneity of some serum glycoproteins in neurodegenerative diseases

Congenital Disorder of Glycosylation IId (CDG-IId) - A New Entity: Clinical Presentation with Dandy-Walker Malformation and Myopathy

Critical Role of Preconceptional Immunization for Protective and Nonpathological Specific Immunity in Murine Neonates

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