A Low Cost Antibody Signal Enhancer Improves Immunolabeling in Cell Culture, Primate Brain and Human Cancer Biopsy
Por:
Flores-Maldonado, Catalina, Estela Albino-Sanchez, M., Rodriguez-Callejas, Juan D., Estrada-Mondragon, Argel, Leon-Galicia, Ismael, Maqueda-Alfaro, Raul, Perez-Cruz, Claudia, Fuchs, Eberhard, Garcia-Carranca, Alejandro, Contreras, Ruben G., Missirlis, Fanis, Rosas-Arellano, Abraham
Publicada:
15 jul 2020
Categoría:
Neuroscience (miscellaneous)
Resumen:
The use of antibodies to identify neuronal receptors, neurotransmitters,
cytoskeletal elements or pathologic protein aggregates, ion channels,
adhesion molecules or other cell-type specific markers, is common
practice in neuroscience. Antibody detection systems are often based on
confocal, epifluorescence or brightfield microscopy. Three types of
technical issues can interfere with immunolabeling: low abundance of the
target protein, low specific affinity of the antibody and/or signal
background sometimes related to tissue fixation. Here, giving tribute to
Professor Miledi's mentorship, we propose the application of an antibody
signal enhancer (ASE) solution based on glycine, hydrogen peroxide and a
detergent mix as a simple, low cost, protocol variation that
significantly and specifically improves the signal to noise ratio during
immunostaining experiments. We describe three new settings in which ASE
improves the detection of a variety of antibodies applied on long-time
stored non-human primate brain sections, cell culture monolayers and on
squamous carcinomas retrieved from cervical cancer patients. The
significant improvement of ASE over optimized immunohistochemical
protocols used in clinical practice (i.e. cancer detection) combined
with its simplicity and low cost makes it an attractive method for
biomedical applications. This article is part of a Special Issue
entitled: Honoring Ricardo Miledi - outstanding neuroscientist of XX-XXI
centuries. (C) 2020 IBRO. Published by Elsevier Ltd. All rights
reserved.
Filiaciones:
Flores-Maldonado, Catalina:
IPN, Dept Fisiol Biofis & Neurociencias, Ctr Invest & Estudios Avanzados, Ciudad De Mexico, Mexico
Estela Albino-Sanchez, M.:
IPN, Dept Biol Celular, Ctr Invest & Estudios Avanzados, Ciudad De Mexico, Mexico
Rodriguez-Callejas, Juan D.:
IPN, Dept Farmacol, Ctr Invest & Estudios Avanzados, Ciudad De Mexico, Mexico
Estrada-Mondragon, Argel:
Linkoping Univ, Dept Clin & Expt Med, Linkoping, Sweden
Leon-Galicia, Ismael:
Univ Nacl Autonoma Mexico, Inst Invest Biomed, Lab Virus & Canc, Unidad Invest Biomed Canc,Dept Biol Mol & Biotecn, Mexico City, DF, Mexico
Secretaria Salud Mexico, Dept Invest Basica, Inst Nacl Cancerol INCan, Mexico City, DF, Mexico
Maqueda-Alfaro, Raul:
IPN, Dept Biol Celular, Ctr Invest & Estudios Avanzados, Ciudad De Mexico, Mexico
Perez-Cruz, Claudia:
IPN, Dept Farmacol, Ctr Invest & Estudios Avanzados, Ciudad De Mexico, Mexico
Fuchs, Eberhard:
Leibniz Inst Primate Res, German Primate Ctr, Gottingen, Germany
Garcia-Carranca, Alejandro:
Univ Nacl Autonoma Mexico, Inst Invest Biomed, Lab Virus & Canc, Unidad Invest Biomed Canc,Dept Biol Mol & Biotecn, Mexico City, DF, Mexico
Secretaria Salud Mexico, Dept Invest Basica, Inst Nacl Cancerol INCan, Mexico City, DF, Mexico
Contreras, Ruben G.:
IPN, Dept Fisiol Biofis & Neurociencias, Ctr Invest & Estudios Avanzados, Ciudad De Mexico, Mexico
Missirlis, Fanis:
IPN, Dept Fisiol Biofis & Neurociencias, Ctr Invest & Estudios Avanzados, Ciudad De Mexico, Mexico
Rosas-Arellano, Abraham:
Univ Nacl Autonoma Mexico, Inst Fisiol Celular, Unidad Imagenol, Apartado Postal 70-242, Ciudad De Mexico 04510, Mexico
|