Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains

Journal of Histochemistry and Cytochemistry

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Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains

JC Adams

Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, Boston 02114.

A procedure is described for intensifying histochemical reactions by amplification of biotinylated sites. This is achieved by deposition of biotinylated tyramine on the tissue through the enzymatic action of horseradish peroxidase (HRP). The amplified biotin sites are subsequently visualized by binding them to avidin, to which a marker is attached. This amplification greatly increases the sensitivity of staining procedures that employ HRP (and/or biotin) in tissue. For neuroanatomical pathway tracing methods, the procedure greatly increases the detectability of the injected tracer. For lectin histochemistry and immunohistochemistry, the amplification requires that the lectin or primary antibody be greatly diluted. This dilution results in less background staining and yet strong signals are produced even when very dilute reagents are used. Alternatively, the amplification permits much shorter incubations in primary antibodies when dilutions are used that would ordinarily be used with conventional bridge techniques. The procedure is also useful for amplifying very weak signals, such as those of immunoreactions in glutaraldehyde-fixed tissue. The amplification procedure, together with the availability of avidin probes labeled with fluorochromes, colloidal gold, or enzyme systems other than HRP, provides a means of greatly increasing the versatility of a variety of histochemical reactions, including those for detecting in situ hybridization probes, in addition to increasing the sensitivity of the reactions.

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M. Jackerott and L.-I. Larsson
Immunocytochemical Localization of the NPY/PYY Y1 Receptor in Enteric Neurons, Endothelial Cells, and Endocrine-like Cells of the Rat Intestinal Tract
J. Histochem. Cytochem., December 1, 1997; 45(12): 1643 - 1650.
[Abstract] [Full Text] [PDF]

R. Malisius, H. Merz, B. Heinz, E. Gafumbegete, B. U. Koch, and A. C. Feller
Constant Detection of CD2, CD3, CD4, and CD5 in Fixed and Paraffin-embedded Tissue Using the Peroxidase-mediated Deposition of Biotin-Tyramide
J. Histochem. Cytochem., December 1, 1997; 45(12): 1665 - 1672.
[Abstract] [Full Text] [PDF]

G. Mayer and M. Bendayan
Biotinyl-Tyramide: A Novel Approach for Electron Microscopic Immunocytochemistry
J. Histochem. Cytochem., November 1, 1997; 45(11): 1449 - 1454.
[Abstract] [Full Text] [PDF]

R. von Wasielewski, M. Mengel, S. Gignac, L. Wilkens, M. Werner, and A. Georgii
Tyramine Amplification Technique in Routine Immunohistochemistry
J. Histochem. Cytochem., November 1, 1997; 45(11): 1455 - 1460.
[Abstract] [Full Text] [PDF]

M. Jackerott and L.-I. Larsson
Immunocytochemical Localization of the NPY/PYY Y1 Receptor in the Developing Pancreas
Endocrinology, November 1, 1997; 138(11): 5013 - 5018.
[Abstract] [Full Text] [PDF]

E. J.M. Speel, F. C.S. Ramaekers, and A. H.N. Hopman
Sensitive Multicolor Fluorescence In Situ Hybridization Using Catalyzed Reporter Deposition (CARD) Amplification
J. Histochem. Cytochem., October 1, 1997; 45(10): 1439 - 1446.
[Abstract] [Full Text] [PDF]

R. R. de Haas, R. P.M. van Gijlswijk, E. B. van der Tol, H. J.M.A.A. Zijlmans, T. Bakker-Schut, J. Bonnet, N. P. Verwoerd, and H. J. Tanke
Platinum Porphyrins as Phosphorescent Label for Time-resolved Microscopy
J. Histochem. Cytochem., September 1, 1997; 45(9): 1279 - 1292.
[Abstract] [Full Text] [PDF]

G. Niedobitek, E. Kremmer, H. Herbst, L. Whitehead, C.W. Dawson, E. Niedobitek, C. von Ostau, N. Rooney, F.A. Grasser, and L.S. Young
Immunohistochemical Detection of the Epstein-Barr Virus-Encoded Latent Membrane Protein 2A in Hodgkin's Disease and Infectious Mononucleosis
Blood, August 15, 1997; 90(4): 1664 - 1672.
[Abstract] [Full Text] [PDF]

P. T. Macechko, L. Krueger, B. Hirsch, and S. L. Erlandsen
Comparison of Immunologic Amplification vs Enzymatic Deposition of Fluorochrome-conjugated Tyramide as Detection Systems for FISH
J. Histochem. Cytochem., March 1, 1997; 45(3): 359 - 364.
[Abstract] [Full Text] [PDF]

B. F. Schmidt, J. Chao, Z. Zhu, R. L. DeBiasio, and G. Fisher
Signal Amplification in the Detection of Single-copy DNA and RNA by Enzyme-catalyzed Deposition (CARD) of the Novel Fluorescent Reporter Substrate Cy3.29-Tyramide
J. Histochem. Cytochem., March 1, 1997; 45(3): 365 - 374.
[Abstract] [Full Text] [PDF]

R. P. M. van Gijlswijk, H. J. M. A. A. Zijlmans, J. Wiegant, M. N. Bobrow, T. J. Erickson, K. E. Adler, H. J. Tanke, and A. K. Raap
Fluorochrome-labeled Tyramides: Use in Immunocytochemistry and Fluorescence In Situ Hybridization
J. Histochem. Cytochem., March 1, 1997; 45(3): 375 - 382.
[Abstract] [Full Text] [PDF]

J. Van heusden, P. de Jong, F. Ramaekers, H. Bruwiere, M. Borgers, and G. Smets
Fluorescein-labeled Tyramide Strongly Enhances the Detection of Low Bromodeoxyuridine Incorporation Levels
J. Histochem. Cytochem., February 1, 1997; 45(2): 315 - 320.
[Abstract] [Full Text] [PDF]

S. B. Bausch and C. Chavkin
Changes in Hippocampal Circuitry after Pilocarpine-Induced Seizures as Revealed by Opioid Receptor Distribution and Activation
J. Neurosci., January 1, 1997; 17(1): 477 - 492.
[Abstract] [Full Text] [PDF]

Z.-Q. Xu, T.-J. Shi, and T. Hokfelt
Expression of galanin and a galanin receptor in several sensory systems and bone anlage of�rat�embryos
PNAS, December 10, 1996; 93(25): 14901 - 14905.
[Abstract] [Full Text] [PDF]

M. M. Daston, M. Bastmeyer, U. Rutishauser, and D. D.�M. O'Leary
Spatially Restricted Increase in Polysialic Acid Enhances Corticospinal Axon Branching Related to Target Recognition and Innervation
J. Neurosci., September 1, 1996; 16(17): 5488 - 5497.
[Abstract] [Full Text] [PDF]

P. Dournaud, Y. Z. Gu, A. Schonbrunn, J. Mazella, G. S. Tannenbaum, and A. Beaudet
Localization of the Somatostatin Receptor SST2A in Rat Brain Using a Specific Anti-Peptide Antibody
J. Neurosci., July 15, 1996; 16(14): 4468 - 4478.
[Abstract] [Full Text] [PDF]

V. Balasingam and V. W. Yong
Attenuation of Astroglial Reactivity by Interleukin-10
J. Neurosci., May 1, 1996; 16(9): 2945 - 2955.
[Abstract] [Full Text] [PDF]

Z Zou, A Anisowicz, M. Hendrix, A Thor, M Neveu, S Sheng, K Rafidi, E Seftor, and R Sager
Maspin, a serpin with tumor-suppressing activity in human mammary epithelial cells
Science, January 28, 1994; 263(5146): 526 - 529.
[Abstract] [PDF]

E. Lazartigues, S. M. Dunlay, A. K. Loihl, P. Sinnayah, J. A. Lang, J. J. Espelund, C. D. Sigmund, and R. L. Davisson
Brain-Selective Overexpression of Angiotensin (AT1) Receptors Causes Enhanced Cardiovascular Sensitivity in Transgenic Mice
Circ. Res., March 22, 2002; 90(5): 617 - 624.
[Abstract] [Full Text] [PDF]