A method for the discrimination of human hemoglobin from animal hemoglobins is described. Hemoglobins of 36 species (human, human fetus, 21 nonhuman primates and 13 non-primate vertebrates) were examined by isoelectric focusing (IEF) in thin layer polyacrylamide gel plates containing an Ampholine gradient of pH3.5-9.5. From the viewpoint of discrimination of human hemoglobin from animal hemoglobins, hemoglobin samples treated with p-chloromercuribenzoate were more effective in IEF than those of untreated. Estimates of the isoelectric points of the major and minor hemoglobins of each species were also described.

View full abstract

LDH isoenzymes in sera from 42 patients with ulcerative colitis were analyzed by means of electrophoretic technique. The abnormal LDH isoenzyme patterns were observed in 4 patients out of them. In the present study, LDH-linked immunoglobulins (Ig) in these 4 patients were examined by immunoelectrophoresis, immunodiffusion and the precipitation method. Then LDH-linked Ig with partial isotypic specifities were identified, i. e., one was LDH-linked IgG (kappa) and the others were LDH-linked IgG (lambda). ALP-linked Ig in ulcerative colitis was identified as IgG in Kano's report. Therefore, the relationship between the isotypic specifity of LDH-linked IgG and ulcerative colitis was speculated.

View full abstract

Clq, Clr and Cls, subcomponents of the first component of complement, have been isolated from human serum in highly purified form by DEAE-Sephacel chromatography and gel filtration through Sepharose 6B and Bio-Gel A 1.5m, and by dialysis against buffers of various ionic strengths. The methods gave excellent reproducibility and recovery. Average yields corresponded to about 16mg, 6mg and 3mg per liter of serum for Clq, Cls and Clr respectively.
The preparations were analyzed by Tris-glycine buffer slab gel electrophoresis, isoelectric focusing gel electrophoresis, and circular dichroism spectra. Approximate molecular weights of activated Clr was 98, 000 and that of Cls was 88, 000.
Analysis by isoelectric gel electrophoresis of Clr and Cls revealed some heterogeneity of isoelectric points. Analysis of circular dichroism spectra of these components showed that Cls mainly had a random coil structure and Clr had substantial amounts of β-structure; also, activated and unactivated forms showed identical spectra. Clr and Cls are activated by very similar mechanisms, but are distinguished by differences of molecular weight, isoelectric point and circular dichroism spectra.

View full abstract