A modification of the amidoblack-TCA-staining for quantitative microspectrometrical determination of proteins in tissue sections.
Conditions are described by which cells and fresh frozen tissues, following fixation with ethanol-ether, and after staining with the amidoblack (AB)-TCA-staining method, show a modified dye/protein ratio of 0.9 moles AB/10(5) g protein compared to 8.5 moles AB/10(5) g protein ( Schauenstein et al. 1980) as in a previously used method. In contrast to the AB-TCA-method, which leads to extremly high and unmeasurable extinctions in tissue sections, staining with the modified AB-TCA- 23st -method with 10 microns tissue sections produces easily measurable extinction values. A correlation of the microspectrometrically determined mean total extinction values of different cell types and nuclei after staining with the tetrazonium method (N ohammer 1978; N ohammer and Desoye 1981) and on the other hand with the AB-TCA 23st -method has been found. The microspectrometrically determined extinctions after AB-TCA 23st -staining can be calculated; an extinction of 0.04248 corresponds to 1 pgm protein.
D000580:Amido Black; D000818:Animals; D001391:Azo Compounds; D002286:Carcinoma, Ehrlich Tumor / Q000032:analysis; D008099:Liver / Q000032:analysis; D008433:Mathematics; D011506:Proteins / Q000032:analysis*; D051381:Rats; D013053:Spectrophotometry; D013194:Staining and Labeling / Q000379:methods*; D014238:Trichloroacetic Acid
D001391:Azo Compounds; D011506:Proteins; D014238:Trichloroacetic Acid; D000580:Amido Black