Suppression of nonspecific binding of avidin-biotin complex (ABC) to proteins electroblotted to nitrocellulose paper.
AuthorsClark RK, Tani Y, Damjanov I
JournalJ Histochem Cytochem
PubMed ID3772081
'Nitrocellulose blots of cell extracts reacted in sequence with biotinylated lectins and horseradish peroxidase-labeled avidin-biotin complex (ABC) often show considerable nonspecific staining of protein bands. Experiments were performed to determine which of the components of the ABC were responsible for this and whether or not the nature and ionic strength ... More
Transmembrane topography of the 100-kDa a subunit (Vph1p) of the yeast vacuolar proton-translocating ATPase.
AuthorsLeng XH, Nishi T, Forgac M
JournalJ Biol Chem
PubMed ID10329659
'The membrane topography of the yeast vacuolar proton-translocating ATPase a subunit (Vph1p) has been investigated using cysteine-scanning mutagenesis. A Cys-less form of Vph1p lacking the seven endogenous cysteines was constructed and shown to have 80% of wild type activity. Single cysteine residues were introduced at 13 sites within the Cys-less ... More
Artifactual detection of biotin on histones by streptavidin.
AuthorsBailey LM, Ivanov RA, Wallace JC, Polyak SW,
JournalAnal Biochem
PubMed ID17920026
'Biotinylation is a recent addition to the list of reported posttranslational modifications made to histones. Holocarboxylase synthetase (HCS) and biotinidase have been implicated as biotinylating enzymes. However, the details of the mechanism and the regulation of biotin transfer on and off histones remains unclear. Here we report that in a ... More
Stimulation of the pentose phosphate pathway and glutathione levels by dehydroascorbate, the oxidized form of vitamin C.
AuthorsPuskas F, Gergely P, Banki K, Perl A
JournalFASEB J
PubMed ID10877828
'Ascorbic acid, or vitamin C, generally functions as an antioxidant by directly reacting with reactive oxygen intermediates and has a vital role in defenses against oxidative stress. However, ascorbic acid also has pro-oxidant properties and may cause apoptosis of lymphoid and myeloid cells. The present study shows that dehydroascorbate, the ... More
Improved immunocytochemical detection of biotinylated probes with Neutralite avidin.
Authorsvan Roy N, Mangelschots K, Speleman F
JournalTrends Genet
PubMed ID8488564
Suppression of endogenous avidin-binding activity in tissues and its relevance to biotin-avidin detection systems.
AuthorsWood GS, Warnke R
JournalJ Histochem Cytochem
PubMed ID7028859
As biotin-avidin systems continue to be developed for applications involving single cells, cell suspensions, and especially tissue sections, the need arises for a method of blocking endogenous avidin-binding activity. One such method is described and its proposed mechanism is discussed. Utilizing this method, endogenous avidin-binding activity was detected and suppressed ... More
Simultaneous inhibition of endogenous avidin-binding activity and peroxidase applicable for the avidin-biotin system using monoclonal antibodies.
AuthorsMatsumoto Y
JournalHistochemistry
PubMed ID2415495
The use of the avidin-biotin technique in immunoperoxidase staining provides a simple and highly sensitive method for detecting the localization of antigens defined by monoclonal antibodies. However, endogenous biotin, which is widely distributed in tissues, often causes non-specific staining by binding to avidin [endogenous avidin-binding activity (EABA)]. Endogenous peroxidase activity ... More
The use of avidin as a probe for the distribution of mitochondrial carboxylases in developing chick retina.
AuthorsRuggiero FP, Sheffield JB
JournalJ Histochem Cytochem
PubMed ID9446824
During development, the inner chick retina progresses from an aerobic to an anaerobic metabolic basis because of the lack of a vascular system. To investigate this process further, we have examined the expression and distribution of mitochondrial carboxylases. Because these enzymes use covalently bound biotin as a co-enzyme, we were ... More
A method for detecting abasic sites in living cells: age-dependent changes in base excision repair.
AuthorsAtamna H, Cheung I, Ames BN
JournalProc Natl Acad Sci U S A
PubMed ID10639140
Apurinic/apyrimidinic (AP) sites are common DNA lesions that arise from spontaneous depurination or by base excision repair (BER) of modified bases. A biotin-containing aldehyde-reactive probe (ARP) [Kubo, K., Ide, H., Wallace, S. S. & Kow, Y. W. (1992) Biochemistry 31, 3703-3708] is used to measure AP sites in living cells. ... More
A new method for simultaneous demonstration of anterograde and retrograde connections in the brain: co-injections of biotinylated dextran amine and the beta subunit of cholera toxin.
In studying reciprocally connected brain networks, it is advantageous to use techniques that allow simultaneous visualization of both efferent and afferent connections from a single injection site. We report on a new technique to achieve this using pressure injections of a mixture of biotinylated dextran amine (BDA) and the beta ... More
CD40 signals apoptosis through FAN-regulated activation of the sphingomyelin-ceramide pathway.
AuthorsSégui B, Andrieu-Abadie N, Adam-Klages S, Meilhac O, Kreder D, Garcia V, Bruno AP, Jaffrézou JP, Salvayre R, Krönke M, Levade T
JournalJ Biol Chem
PubMed ID10601289
The possibility that the sphingomyelin (SM)-ceramide pathway is activated by CD40, a transmembrane glycoprotein belonging to the tumor necrosis factor receptor superfamily and that plays a critical role in the regulation of immune responses has been investigated. We demonstrate that incubation of Epstein-Barr virus-transformed lymphoid cells with an anti-CD40 antibody ... More
Comparative morphology of three types of projection-identified pyramidal neurons in the superficial layers of cat visual cortex.
AuthorsMatsubara JA, Chase R, Thejomayen M
JournalJ Comp Neurol
PubMed ID8866848
The morphology and dendritic organization of corticocortical neurons in the superficial layers of area 18 that project to area 17 were studied by intracellular injection of lucifer yellow in the fixed-slice preparation. This corticocortical population contains primarily standard pyramidal cells, but occasional nonpyramidal, modified, fusiform, star, and inverted pyramidal cells ... More
The human AC133 hematopoietic stem cell antigen is also expressed in epithelial cells and targeted to plasma membrane protrusions.
AuthorsCorbeil D, Röper K, Hellwig A, Tavian M, Miraglia S, Watt SM, Simmons PJ, Peault B, Buck DW, Huttner WB
JournalJ Biol Chem
PubMed ID10681530
The human AC133 antigen and mouse prominin are structurally related plasma membrane proteins. However, their tissue distribution is distinct, with the AC133 antigen being found on hematopoietic stem and progenitor cells and prominin on various epithelial cells. To determine whether the human AC133 antigen and mouse prominin are orthologues or ... More