The use of avidin-biotin interaction in immunoenzymatic techniques.
AuthorsGuesdon JL, Ternynck T, Avrameas S
JournalJ Histochem Cytochem
PubMed ID90074
'Biotin was covalently attached to antibodies, antigens and enzymes, and the effects of this labeling on the antigen and antibody binding capacity and on enzymatic activity were tested. Based on avidin-biotin interaction, the labeled proteins were used in quantitative enzyme-immunoassay and enzyme-immunohistochemical staining procedures. Two procedures were developed. In the ... More
A rapid and sensitive non-competitive avidin-biotin assay for lactoferrin.
AuthorsVilja P, Krohn K, Tuohimaa P
JournalJ Immunol Methods
PubMed ID3881534
'We have developed an avidin-biotin assay for the detection of lactoferrin in human seminal plasma. We compared 5 modifications of this assay, and found the non-competitive avidin-biotin assay (NABA) to be the most sensitive. The detection limit of 3-step NABA was 0.2 or 0.1 ng lactoferrin/ml, depending whether avidin-biotin-peroxidase complex ... More
Conjugated avidin binds to mast cell granules.
AuthorsTharp MD, Seelig LL, Tigelaar RE, Bergstresser PR
JournalJ Histochem Cytochem
PubMed ID2578142
'The glycoprotein, avidin, conjugated either to the enzyme horseradish peroxidase, or to the fluorochrome dyes, fluorescein or rhodamine, identifies the granules of mast cells in both tissues and cell suspensions. In the absence of prior fixation, mast cells were not identified with conjugated avidin; however, granules released from these cells ... More
3-(N-Maleimido-propionyl)biocytin: a versatile thiol-specific biotinylating reagent.
AuthorsBayer EA, Zalis MG, Wilchek M
JournalAnal Biochem
PubMed ID3935007
'A biotin-containing, thiol-specific reagent, 3-(N-maleimido-propionyl) biocytin (MPB), was synthesized and used to biotinylate various proteins via native or artificially induced sulfhydryl groups. In combination with appropriate avidin- or streptavidin-conjugated markers (i.e., fluorescent, enzyme-conjugated, electron-dense, etc.), MPB essentially constitutes a universal, multipurpose, thiol-specific probe. The reagent could be used to detect ... More
The SA/rABC technique: a new ABC procedure for detection of antigens at increased sensitivity.
AuthorsGrumbach IM, Veh RW
JournalJ Histochem Cytochem
PubMed ID7822761
'Since its introduction, the avidin-biotin-peroxidase (ABC) complex has become an invaluable detection system for a wide variety of bioanalytical applications. In these techniques, avidin and biotin-peroxidase are mixed at a pre-determined ratio so that the soluble ABC complex retains biotin binding sites. Consequently, the complex contains an excess of avidin ... More
Enhanced chemiluminescent method for the detection of DNA dot-hybridization assays.
AuthorsMatthews JA, Batki A, Hynds C, Kricka LJ
JournalAnal Biochem
PubMed ID4091282
A simple enhanced chemiluminescent procedure for the quantitation of DNA hybridization to dot blots is described. The method utilizes DNA probes labeled with biotin, which are detected using a biotinylated streptavidin-horseradish peroxidase complex. The peroxidase enzyme then takes part in an enhanced chemiluminescent reaction with luminol, peroxide, and an enhancer. ... More
AuthorsCattoretti G, Berti E, Schiró R, D'Amato L, Valeggio C, Rilke F
JournalHistochem J
PubMed ID2455699
A considerable intensification of the avidin-biotin-peroxidase complex staining system (ABC) was obtained by sequentially overlaying the sections to be immunostained with an avidin-rich and a biotin-rich complex. Each sequential addition contributed to the deposition of horseradish peroxidase on the immunostained site and allowed the subsequent binding of a complementary complex. ... More
Visual detection of granulocyte surface antigens using the avidin-biotin complex.
AuthorsHenke M, Yonemoto LM, Lazar GS, Gaidulis L, Hecht T, Santos S, Blume KG
JournalJ Histochem Cytochem
PubMed ID6376617
A visual test for detection of granulocyte surface markers using the avidin-biotin complex (ABC) has been developed. That this assay is highly specific, reproducible, and sensitive was determined by studying the expression of HLA antigens on granulocytes with monoclonal antibodies. Further, using granulocyte specific alloantisera, the results of the ABC ... More
Biotinylation: a simple method for labelling complement component C8 with preservation of functional activity.
AuthorsBhakdi S, Roth M, Hugo F
JournalJ Immunol Methods
PubMed ID2754261
Biotinylation of human C8 with the water-soluble biotin derivative biotinylamidohexanoic acid, N-hydroxysulfosuccinimide ester is an excellent method for labelling this terminal complement component with preservation of its functional activity. The biotinylated product can be detected both in native form and also following its incorporation into the terminal complement complexes. Detection ... More
Selective staining of platelet glycoproteins using two-dimensional O'Farrell gel electrophoresis and avidin-biotin-conjugated lectins.
AuthorsKehrel B, Kokott R, Balleisen L, Stenzinger W, Clemetson KJ, van de Loo J
JournalThromb Haemost
PubMed ID2451305
A time-saving and sensitive high resolution method for the analysis and identification of platelet glycoproteins using nonradioactive compounds has been developed. Platelet proteins (50 micrograms) of normal subjects were separated by isoelectric focusing and SDS-PAGE. Proteins were either stained with silver or electroblotted onto nitrocellulose. Nitrocellulose blots were treated with ... More
Versatility of biotin-labeled lectins and avidin-biotin-peroxidase complex for localization of carbohydrate in tissue sections.
AuthorsHsu SM, Raine L
JournalJ Histochem Cytochem
PubMed ID7037937
Carbohydrate residues in tissue can be localized by a very sensitive, specific, and simple technique, utilizing a biotin-labeled lectin followed by an avidin-biotin-peroxidase complex (ABC). Distribution of peanut receptors in benign and malignant tissues was found to be quite different. Normal colonic mucosa occasionally shows small dot staining in the ... More
Horseradish peroxidase: a modern view of a classic enzyme.
AuthorsVeitch NC,
JournalPhytochemistry
PubMed ID14751298
Horseradish peroxidase is an important heme-containing enzyme that has been studied for more than a century. In recent years new information has become available on the three-dimensional structure of the enzyme and its catalytic intermediates, mechanisms of catalysis and the function of specific amino acid residues. Site-directed mutagenesis and directed ... More
Ultrastructural localization of lectin receptors on the surface of the rat retinal pigment epithelium. Decreased sensitivity of the avidin-biotin method due to cell surface charge.
AuthorsPino RM
JournalJ Histochem Cytochem
PubMed ID6747275
Monosaccharides on the apical processes of the retinal pigment epithelium were examined using lectin-affinity cytochemical methods. Lectin receptor sugars were localized with lectin-horseradish peroxidase (HRP) and lectin-ferritin conjugates as well as with biotinylated lectins, avidin, and biotinylated HRP. In contrast, only wheat germ agglutinin (WGA) receptors were identified with biotinylated ... More
Cellular origin of fibronectin in interspecies hybrid kidneys.
AuthorsSariola H, Kuusela P, Ekblom P
JournalJ Cell Biol
PubMed ID6389571
The cellular origin of fibronectin in the kidney was studied in three experimental models. Immunohistochemical techniques that use cross-reacting or species-specific antibodies against mouse or chicken fibronectin were employed. In the first model studied, initially avascular mouse kidneys cultured on avian chorioallantoic membranes differentiate into epithelial kidney tubules and become ... More
Selective labeling of sulfhydryls and disulfides on blot transfers using avidin-biotin technology: studies on purified proteins and erythrocyte membranes.
AuthorsBayer EA, Safars M, Wilchek M
JournalAnal Biochem
PubMed ID2437828
Various strategies for the use of 3-(N-maleimido-propionyl) biocytin (MPB) as a general label for distinguishing between protein sulfhydryls and disulfides on blot transfers are presented. In the first approach, endogenous SH groups in proteins were labeled directly with MPB. For disulfide staining, endogenous sulfhydryls were blocked with N-ethylmaleimide, disulfides were ... More
The cell surface of a restrictive fenestrated endothelium. I. Distribution of lectin-receptor monosaccharides on the choriocapillaris.
AuthorsPino RM
JournalCell Tissue Res
PubMed ID3943119
The choriocapillaris is one example of a capillary bed lined by a fenestrated endothelium that is restrictive to exogenous tracers and endogenous plasma proteins. In this study we have examined the distribution of cell-surface monosaccharides utilizing biotinylated lectin-avidin ferritin cytochemistry. Receptors for wheat germ agglutinin were localized to the plasmalemma ... More