An immunofluorescent method for a specific demonstration of granulocytes and some of their proteins (ECP and CCP)

Using p-benzoquinone as a fixative the non-specific fluorescence of granulocytes and especially the eosinophils is removed for both FITC and TRITC. In this way we have been able to detect the eosinophil cationic protein (ECP) and the chymotrypsin-like cationic protein (CCP) in human lung tissue and by double immunofluorescent labelling shown that these two proteins very likely are related to eosinophils respectively the neutrophils.     

An alternate method for demonstration of bisphosphoglyceromutase (DPGM) on starch gels

The phosphatase activity of bisphosphoglyceromutase (DPGM) was used to determine the phenotypes of the enzyme. DPGM was polymorphic in four Alaskan ethnic groups.     

An argyrophil III method for the demonstration of micro- and oligodendroglia

Micro- and oligodendroglia, plasma and nucleoli of nerve cells, capillary wall and nuclei of astrocytes become visible when sections of formol fixed human brain are immersed, without any previous treatment, into a physical developer of pH 10.5. The staining is inhibited by the catalytic activity of the tissue elements involved. By means of pretreatments with 1% performic acid and 30% sodium rhodanide dissolved in 0.4% sodium hydroxide, the catalytic activity in the unwanted tissue elements is suppressed, and this results in an elective demonstration of micro- and oligodendroglia. Reducing groups of the tissue or any kind of performed nuclei play no role in this silver staining.     

A specific histochemical method for the demonstration of the activity of pancreatic lipase

Synopsis Emulsified long-chain triglyceride, a specific substrate for the enzyme pancreatic lipase (glycerol-ester hydrolase, EC 3.1.1.3), has been used in a modification of the Gomori technique for the demonstration of lipase. In the range of tissues examined (pancreas, testis, cardiac stomach and liver), true pancreatic lipase activity was revealed only in pancreatic tissue, by contrast with results obtained with less specific methods.     

An argyrophil III method for the demonstration of fibrous neuroglia

The catalytic activity of the fibrous astrocytes in the reaction of silver ions with dissolved reducing molecules is considerably increased by ethylation and a subsequent treatment with NaJO3. As a consequence, in a special physical developer they produce metallic silver at a considerably higher rate than do other tissue elements, making possible their selective demonstration. In freshly fixed materials myelin too is silverized. This can be avoided by means of pretreatments with performic acid and iodine.     

An Alcian blue (pH 2.5)-PAS-keratin immunoperoxidase method for the simultaneous demonstration of keratin and neutral and acidic mucosubstances

A method is proposed for the simultaneous staining of neutral and acidic, periodate reactive and nonperiodate reactive mucosubstances, glycogen and keratin in paraffin sections. Briefly, sections are stained by the Alcian blue (pH 2.5)-PAS method, followed by a peroxidase-antiperoxidase immunohistochemical stain for keratin. The proposed method modifies an existing method, and expands the range of polysaccharides and mucosubstances which may be demonstrated. The proposed method is easily performed within a single working day and promises to be of value in surgical pathology as well as in studies of bronchial carcinogenesis.     

A sensitive and specific method for quantitative estimation of carbamylation in proteins

A simple, rapid, reproducible, and specific micromethod for the estimation of carbamylation of proteins is described. The method is based on permanganate oxidation of radioactive carbamyl derivatives of protein to form urea and on the specific decomposition of urea by urease.     

An updated nomenclature for keratin-associated proteins (KAPs)

Most protein in hair and wool is of two broad types: keratin intermediate filament-forming proteins (commonly known as keratins) and keratin-associated proteins (KAPs). Keratin nomenclature was reviewed in 2006, but the KAP nomenclature has not been revised since 1993. Recently there has been an increase in the number of KAP genes (KRTAPs) identified in humans and other species, and increasingly reports of variation in these genes. We therefore propose that an updated naming system is needed to accommodate the complexity of the KAPs. It is proposed that the system is founded in the previous nomenclature, but with the abbreviation sp-KAPm-nL*x for KAP proteins and sp-KRTAPm-n(p/L)*x for KAP genes. In this system sp is a unique letter-based code for different species as described by the protein knowledge-based UniProt. m is a number identifying the gene or protein family, n is a constituent member of that family, p signifies a pseudogene if present, L if present signifies like and refers to a temporary place-holder until the family is confirmed and x signifies a genetic variant or allele. We support the use of non-italicised text for the proteins and italicised text for the genes. This nomenclature is not that different to the existing system, but it includes species information and also describes genetic variation if identified, and hence is more informative. For example, GenBank sequence JN091630 would historically have been named KRTAP7-1 for the gene and KAP7-1 for the protein, but with the proposed nomenclature would be SHEEP-KRTAP7-1*A and SHEEP-KAP7-1*A for the gene and protein respectively. This nomenclature will facilitate more efficient storage and retrieval of data and define a common language for the KAP proteins and genes from all mammalian species.     

Use of electroblotting as a method for preparing samples of proteins and their fragments for microsequencing

Gel electrophoresis in the presence of sodium dodecyl sulphate followed by electroblotting was employed in sample preparation for microsequencing proteins and protein fragments. Three types of solid supports were compared: glass fiber filters modified by aminopropyltriethoxysilane or covered with polybrene, and polyvinylidenedifluoride membranes. N-Terminal amino acid sequences of several proteins (Mr 14-140 kDA were determined on a gas-phase sequencer with the standard programme; 20-200 pmoles of the protein can be assayed by this method.     

An alternate method for demonstration of erythrocytic aminotransferases on starch gels

A two-stage procedure using MTT tetrazolium for the demonstration of aminotransferases (GPT and GOT) either singly or together was developed. Identification of phenotypes was unequivocal in over 99% of the individuals studied.     

Unfixed material embedded in a methacrylate resin (Technovit 7100) for immunofluorescent staining

Wheat grains were embedded with or without fixation in Technovit 7100 or in paraffin. This enabled us to produce 4 microns sections for immunofluorescent staining to see whether serum of patients with baker's asthma contained IgG antibodies against wheat grains. Embedding without fixation in Technovit 7100 appeared to be suitable for immunofluorescent staining and gave superior results to protocols requiring fixation.