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.

Volume 40, Issue 10, pp. 1457-1463, 10/01/1992
Copyright © 1992 by The Histochemical Society

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				I. C. Maier, K. Baumann, M. Thallmair, O. Weinmann, J. Scholl, and M. E. Schwab Constraint-Induced Movement Therapy in the Adult Rat after Unilateral Corticospinal Tract Injury J. Neurosci., September 17, 2008; 28(38): 9386 - 9403. [Abstract] [Full Text] [PDF]

				T. Furuta, E. Timofeeva, K. Nakamura, K. Okamoto-Furuta, M. Togo, T. Kaneko, and M. Deschenes Inhibitory Gating of Vibrissal Inputs in the Brainstem J. Neurosci., February 20, 2008; 28(8): 1789 - 1797. [Abstract] [Full Text] [PDF]

				E. L. Dimitrov, M. R. DeJoseph, M. S. Brownfield, and J. H. Urban Involvement of Neuropeptide Y Y1 Receptors in the Regulation of Neuroendocrine Corticotropin-Releasing Hormone Neuronal Activity Endocrinology, August 1, 2007; 148(8): 3666 - 3673. [Abstract] [Full Text] [PDF]

				S. Bensing, S. O. Fetissov, J. Mulder, J. Perheentupa, J. Gustafsson, E. S. Husebye, M. Oscarson, O. Ekwall, P. A. Crock, T. Hokfelt, et al. Pituitary autoantibodies in autoimmune polyendocrine syndrome type 1 PNAS, January 16, 2007; 104(3): 949 - 954. [Abstract] [Full Text] [PDF]

				P. Bergman, L. Johansson, H. Wan, A. Jones, R. L. Gallo, G. H. Gudmundsson, T. Hokfelt, A.-B. Jonsson, and B. Agerberth Induction of the Antimicrobial Peptide CRAMP in the Blood-Brain Barrier and Meninges after Meningococcal Infection Infect. Immun., December 1, 2006; 74(12): 6982 - 6991. [Abstract] [Full Text] [PDF]

				P. Sinnayah, E. Lazartigues, K. Sakai, R. V. Sharma, C. D. Sigmund, and R. L. Davisson Genetic Ablation of Angiotensinogen in the Subfornical Organ of the Brain Prevents the Central Angiotensinergic Pressor Response Circ. Res., November 10, 2006; 99(10): 1125 - 1131. [Abstract] [Full Text] [PDF]

				H. P. Nguyen, P. Kobbe, H. Rahne, T. Worpel, B. Jager, M. Stephan, R. Pabst, C. Holzmann, O. Riess, H. Korr, et al. Behavioral abnormalities precede neuropathological markers in rats transgenic for Huntington's disease Hum. Mol. Genet., November 1, 2006; 15(21): 3177 - 3194. [Abstract] [Full Text] [PDF]

				J. H. Urban, R. J. Leitermann, M. R. DeJoseph, S. J. Somponpun, M. L. Wolak, and C. D. Sladek Influence of Dehydration on the Expression of Neuropeptide Y Y1 Receptors in Hypothalamic Magnocellular Neurons Endocrinology, September 1, 2006; 147(9): 4122 - 4131. [Abstract] [Full Text] [PDF]

				J Svensson, M Diez, J Engel, C Wass, A Tivesten, J-O Jansson, O Isaksson, T Archer, T Hokfelt, and C Ohlsson Endocrine, liver-derived IGF-I is of importance for spatial learning and memory in old mice. J. Endocrinol., June 1, 2006; 189(3): 617 - 627. [Abstract] [Full Text] [PDF]

				A. Svensson, J. Kaim, C. Mallard, A. Olsson, E. Brodin, T. Hokfelt, and K. Eriksson Neurokinin 1 Receptor Signaling Affects the Local Innate Immune Defense against Genital Herpes Virus Infection J. Immunol., November 15, 2005; 175(10): 6802 - 6811. [Abstract] [Full Text] [PDF]

				A. F. Zehnder, J. C. Adams, P. A. Santi, A. G. Kristiansen, C. Wacharasindhu, S. Mann, R. Kalluri, M. C. Gregory, C. E. Kashtan, and S. N. Merchant Distribution of Type IV Collagen in the Cochlea in Alport Syndrome Arch Otolaryngol Head Neck Surg, November 1, 2005; 131(11): 1007 - 1013. [Abstract] [Full Text] [PDF]

				J.-B. Shin, D. Adams, M. Paukert, M. Siba, S. Sidi, M. Levin, P. G. Gillespie, and S. Grunder Xenopus TRPN1 (NOMPC) localizes to microtubule-based cilia in epithelial cells, including inner-ear hair cells PNAS, August 30, 2005; 102(35): 12572 - 12577. [Abstract] [Full Text] [PDF]

				J. A. Ramos-Vara Technical Aspects of Immunohistochemistry Vet. Pathol., July 1, 2005; 42(4): 405 - 426. [Abstract] [Full Text] [PDF]

				L. Johansson, A. Rytkonen, H. Wan, P. Bergman, L. Plant, B. Agerberth, T. Hokfelt, and A.-B. Jonsson Human-Like Immune Responses in CD46 Transgenic Mice J. Immunol., July 1, 2005; 175(1): 433 - 440. [Abstract] [Full Text] [PDF]

				A.-M. Bao, A. Hestiantoro, E. J. W. Van Someren, D. F. Swaab, and J.-N. Zhou Colocalization of corticotropin-releasing hormone and oestrogen receptor-{alpha} in the paraventricular nucleus of the hypothalamus in mood disorders Brain, June 1, 2005; 128(6): 1301 - 1313. [Abstract] [Full Text] [PDF]

				E. Hrabovszky, G. Wittmann, G. F. Turi, Z. Liposits, and C. Fekete Hypophysiotropic Thyrotropin-Releasing Hormone and Corticotropin-Releasing Hormone Neurons of the Rat Contain Vesicular Glutamate Transporter-2 Endocrinology, January 1, 2005; 146(1): 341 - 347. [Abstract] [Full Text] [PDF]

				U. C. Kopp, M. Z. Cicha, K. Nakamura, R. M. Nusing, L. A. Smith, and T. Hokfelt Activation of EP4 receptors contributes to prostaglandin E2-mediated stimulation of renal sensory nerves Am J Physiol Renal Physiol, December 1, 2004; 287(6): F1269 - F1282. [Abstract] [Full Text] [PDF]

				S. Threlfell, S. J. Cragg, I. Kallo, G. F. Turi, C. W. Coen, and S. A. Greenfield Histamine H3 Receptors Inhibit Serotonin Release in Substantia Nigra Pars Reticulata J. Neurosci., October 6, 2004; 24(40): 8704 - 8710. [Abstract] [Full Text] [PDF]

				J. Oldekamp, N. Kraemer, G. Alvarez-Bolado, and T. Skutella bHLH Gene Expression in the Emx2-deficient Dentate Gyrus Reveals Defective Granule Cells and Absence of Migrating Precursors Cereb Cortex, September 1, 2004; 14(9): 1045 - 1058. [Abstract] [Full Text] [PDF]

				R. Fossmark, T. C. Martinsen, K. E. Bakkelund, S. Kawase, and H. L. Waldum ECL-Cell Derived Gastric Cancer in Male Cotton Rats Dosed with the H2-Blocker Loxtidine Cancer Res., May 15, 2004; 64(10): 3687 - 3693. [Abstract] [Full Text] [PDF]

				R. Nardacci, I. Falciatori, S. Moreno, and S. Stefanini Immunohistochemical Localization of Peroxisomal Enzymes During Rat Embryonic Development J. Histochem. Cytochem., April 1, 2004; 52(4): 423 - 436. [Abstract] [Full Text] [PDF]

				N. Shigemura, R. Ohta, Y. Kusakabe, H. Miura, A. Hino, K. Koyano, K. Nakashima, and Y. Ninomiya Leptin Modulates Behavioral Responses to Sweet Substances by Influencing Peripheral Taste Structures Endocrinology, February 1, 2004; 145(2): 839 - 847. [Abstract] [Full Text] [PDF]

				K. Jin, A. L. Peel, X. O. Mao, L. Xie, B. A. Cottrell, D. C. Henshall, and D. A. Greenberg Increased hippocampal neurogenesis in Alzheimer's disease PNAS, January 6, 2004; 101(1): 343 - 347. [Abstract] [Full Text] [PDF]

				T. Yoshitake, F.-H. Wang, E. Kuteeva, K. Holmberg, M. Yamaguchi, J. N. Crawley, R. Steiner, T. Bartfai, S. O. Ogren, T. Hokfelt, et al. Enhanced hippocampal noradrenaline and serotonin release in galanin-overexpressing mice after repeated forced swimming test PNAS, January 6, 2004; 101(1): 354 - 359. [Abstract] [Full Text] [PDF]

				T. C. Martinsen, S. Kawase, R. Hakanson, S. H. Torp, R. Fossmark, G. Qvigstad, A. K. Sandvik, and H. L. Waldum Spontaneous ECL cell carcinomas in cotton rats: natural course and prevention by a gastrin receptor antagonist Carcinogenesis, December 1, 2003; 24(12): 1887 - 1896. [Abstract] [Full Text] [PDF]

				G. F. Turi, Z. Liposits, S. M. Moenter, C. Fekete, and E. Hrabovszky Origin of Neuropeptide Y-Containing Afferents to Gonadotropin-Releasing Hormone Neurons in Male Mice Endocrinology, November 1, 2003; 144(11): 4967 - 4974. [Abstract] [Full Text] [PDF]

				A Iannaccone, D K Breuer, X F Wang, S F Kuo, E M Normando, E Filippova, A Baldi, S Hiriyanna, C B MacDonald, F Baldi, et al. Clinical and immunohistochemical evidence for an X linked retinitis pigmentosa syndrome with recurrent infections and hearing loss in association with an RPGR mutation J. Med. Genet., November 1, 2003; 40(11): e118 - 118. [Full Text] [PDF]

				T. Uchihara, A. Nakamura, H. Nakayama, K. Arima, N. Ishizuka, H. Mori, and S. Mizushima Triple Immunofluorolabeling with Two Rabbit Polyclonal Antibodies and a Mouse Monoclonal Antibody Allowing Three-dimensional Analysis of Cotton Wool Plaques in Alzheimer Disease J. Histochem. Cytochem., September 1, 2003; 51(9): 1201 - 1206. [Abstract] [Full Text] [PDF]

				N. Vrang, C. B. Phifer, M. M. Corkern, and H.-R. Berthoud Gastric distension induces c-Fos in medullary GLP-1/2-containing neurons Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2003; 285(2): R470 - R478. [Abstract] [Full Text] [PDF]

				L. Johansson, A. Rytkonen, P. Bergman, B. Albiger, H. Kallstrom, T. Hokfelt, B. Agerberth, R. Cattaneo, and A.-B. Jonsson CD46 in Meningococcal Disease Science, July 18, 2003; 301(5631): 373 - 375. [Abstract] [Full Text] [PDF]

				A. Schad, H. D. Fahimi, A. Volkl, and E. Baumgart Expression of Catalase mRNA and Protein in Adult Rat Brain: Detection by Nonradioactive In Situ Hybridization with Signal Amplification by Catalyzed Reporter Deposition (ISH-CARD) and Immunohistochemistry (IHC)/Immunofluorescence (IF) J. Histochem. Cytochem., June 1, 2003; 51(6): 751 - 760. [Abstract] [Full Text] [PDF]

				O. Beltcheva, S. Kontusaari, S. Fetissov, H. Putaala, P. Kilpelainen, T. Hokfelt, and K. Tryggvason Alternatively Used Promoters and Distinct Elements Direct Tissue-Specific Expression of Nephrin J. Am. Soc. Nephrol., February 1, 2003; 14(2): 352 - 358. [Abstract] [Full Text] [PDF]

				S. O. Fetissov, J. Hallman, L. Oreland, B. af Klinteberg, E. Grenback, A.-L. Hulting, and T. Hokfelt Autoantibodies against alpha -MSH, ACTH, and LHRH in anorexia and bulimia nervosa patients PNAS, December 24, 2002; 99(26): 17155 - 17160. [Abstract] [Full Text] [PDF]

				M. C. Zimmerman, E. Lazartigues, J. A. Lang, P. Sinnayah, I. M. Ahmad, D. R. Spitz, and R. L. Davisson Superoxide Mediates the Actions of Angiotensin II in the Central Nervous System Circ. Res., November 29, 2002; 91(11): 1038 - 1045. [Abstract] [Full Text] [PDF]

				I. Brouns, L. Van Nassauw, J. Van Genechten, M. Majewski, D. W. Scheuermann, J.-P. Timmermans, and D. Adriaensen Triple Immunofluorescence Staining with Antibodies Raised in the Same Species to Study the Complex Innervation Pattern of Intrapulmonary Chemoreceptors J. Histochem. Cytochem., April 1, 2002; 50(4): 575 - 582. [Abstract] [Full Text] [PDF]

				I. B. BUCHWALOW, T. PODZUWEIT, W. BOCKER, V. E. SAMOILOVA, S. THOMAS, M. WELLNER, H. A. BABA, H. ROBENEK, J. SCHNEKENBURGER, and M. M. LERCH Vascular smooth muscle and nitric oxide synthase FASEB J, April 1, 2002; 16(6): 500 - 508. [Abstract] [Full Text] [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]

				S. L. Barrans, I. Carter, R. G. Owen, F. E. Davies, R. D. Patmore, A. P. Haynes, G. J. Morgan, and A. S. Jack Germinal center phenotype and bcl-2 expression combined with the International Prognostic Index improves patient risk stratification in diffuse large B-cell lymphoma Blood, February 15, 2002; 99(4): 1136 - 1143. [Abstract] [Full Text] [PDF]

				X. Z. Liu, X. J. Xia, J. Adams, Z. Y. Chen, K. O. Welch, M. Tekin, X. M. Ouyang, A. Kristiansen, A. Pandya, T. Balkany, et al. Mutations in GJA1 (connexin 43) are associated with non-syndromic autosomal recessive deafness Hum. Mol. Genet., December 1, 2001; 10(25): 2945 - 2951. [Abstract] [Full Text] [PDF]

				N. G. Robertson, B. L. Resendes, J. S. Lin, C. Lee, J. C. Aster, J. C. Adams, and C. C. Morton Inner ear localization of mRNA and protein products of COCH, mutated in the sensorineural deafness and vestibular disorder, DFNA9 Hum. Mol. Genet., October 1, 2001; 10(22): 2493 - 2500. [Abstract] [Full Text] [PDF]

				Q. Liu, K. G. Hamil, P. Sivashanmugam, G. Grossman, R. Soundararajan, A. J. Rao, R. T. Richardson, Y.-L. Zhang, M. G. O'Rand, P. Petrusz, et al. Primate Epididymis-Specific Proteins: Characterization of ESC42, a Novel Protein Containing a Trefoil-Like Motif in Monkey and Human Endocrinology, October 1, 2001; 142(10): 4529 - 4539. [Abstract] [Full Text] [PDF]

				L. M. Gjerdrum, I. Lielpetere, L. M. Rasmussen, K. Bendix, and S. Hamilton-Dutoit Laser-Assisted Microdissection of Membrane-Mounted Paraffin Sections for Polymerase Chain Reaction Analysis: Identification of Cell Populations Using Immunohistochemistry and in Situ Hybridization J. Mol. Diagn., August 1, 2001; 3(3): 105 - 110. [Abstract] [Full Text] [PDF]

				A. L. Peel, R. V. Rao, B. A. Cottrell, M. R. Hayden, L. M. Ellerby, and D. E. Bredesen Double-stranded RNA-dependent protein kinase, PKR, binds preferentially to Huntington's disease (HD) transcripts and is activated in HD tissue Hum. Mol. Genet., July 1, 2001; 10(15): 1531 - 1538. [Abstract] [Full Text] [PDF]

				U. C. Kopp, M. Z. Cicha, L. A. Smith, and T. Hokfelt Nitric oxide modulates renal sensory nerve fibers by mechanisms related to substance P receptor activation Am J Physiol Regulatory Integrative Comp Physiol, July 1, 2001; 281(1): R279 - R290. [Abstract] [Full Text] [PDF]

				L. C. Sincich and G. G. Blasdel Oriented Axon Projections in Primary Visual Cortex of the Monkey J. Neurosci., June 15, 2001; 21(12): 4416 - 4426. [Abstract] [Full Text] [PDF]

				F. P. M. Kruijver, A. Fernández-Guasti, M. Fodor, E. M. Kraan, and D. F. Swaab Sex Differences in Androgen Receptors of the Human Mamillary Bodies Are Related to Endocrine Status Rather Than to Sexual Orientation or Transsexuality J. Clin. Endocrinol. Metab., February 1, 2001; 86(2): 818 - 827. [Abstract] [Full Text]

				C. Fekete, E. Mihaly, L.-G. Luo, J. Kelly, J. T. Clausen, Q. Mao, W. M. Rand, L. G. Moss, M. Kuhar, C. H. Emerson, et al. Association of Cocaine- and Amphetamine-Regulated Transcript-Immunoreactive Elements with Thyrotropin-Releasing Hormone-Synthesizing Neurons in the Hypothalamic Paraventricular Nucleus and Its Role in the Regulation of the Hypothalamic-Pituitary-Thyroid Axis during Fasting J. Neurosci., December 15, 2000; 20(24): 9224 - 9234. [Abstract] [Full Text] [PDF]

				J. F. Breininger and D. G. Baskin Fluorescence In Situ Hybridization of Scarce Leptin Receptor mRNA using the Enzyme-Labeled Fluorescent Substrate Method and Tyramide Signal Amplification J. Histochem. Cytochem., December 1, 2000; 48(12): 1593 - 1600. [Abstract] [Full Text]

				B. Serafini, S. Columba-Cabezas, F. Di Rosa, and F. Aloisi Intracerebral Recruitment and Maturation of Dendritic Cells in the Onset and Progression of Experimental Autoimmune Encephalomyelitis Am. J. Pathol., December 1, 2000; 157(6): 1991 - 2002. [Abstract] [Full Text] [PDF]

				M. Gaster, J. Franch, P. Staehr, H. Beck-Nielsen, T. Smith, and H. D. Schroder Induction of GLUT-1 protein in adult human skeletal muscle fibers Am J Physiol Endocrinol Metab, November 1, 2000; 279(5): E1191 - E1195. [Abstract] [Full Text] [PDF]

				G. M. Portela-Gomes, A. Lukinius, and L. Grimelius Synaptic Vesicle Protein 2, A New Neuroendocrine Cell Marker Am. J. Pathol., October 1, 2000; 157(4): 1299 - 1309. [Abstract] [Full Text] [PDF]

				M. Gaster, A. Handberg, H. Beck-Nielsen, and H. D. Schroder Glucose transporter expression in human skeletal muscle fibers Am J Physiol Endocrinol Metab, September 1, 2000; 279(3): E529 - E538. [Abstract] [Full Text] [PDF]

				A. Köhler, B. Lauritzen, and C. J.F. Van Noorden Signal Amplification in Immunohistochemistry at the Light Microscopic Level Using Biotinylated Tyramide and Nanogold-Silver Staining J. Histochem. Cytochem., July 1, 2000; 48(7): 933 - 942. [Abstract] [Full Text]

				L. G. Kömüves, A. Feren, A. L. Jones, and E. Fodor Expression of Epidermal Growth Factor and Its Receptor in Cirrhotic Liver Disease J. Histochem. Cytochem., June 1, 2000; 48(6): 821 - 830. [Abstract] [Full Text]

				S. Schulz, S. U. Pauli, S. Schulz, M. Händel, K. Dietzmann, R. Firsching, and V. Höllt Immunohistochemical Determination of Five Somatostatin Receptors in Meningioma Reveals Frequent Overexpression of Somatostatin Receptor Subtype sst2A Clin. Cancer Res., May 1, 2000; 6(5): 1865 - 1874. [Abstract] [Full Text]

				G. Mayer, R. D. Leone, J. F. Hainfeld, and M. Bendayan Introduction of a Novel HRP Substrate-Nanogold Probe for Signal Amplification in Immunocytochemistry J. Histochem. Cytochem., April 1, 2000; 48(4): 461 - 470. [Abstract] [Full Text]

				C. Broberger, K. Holmberg, M. J. Kuhar, and T. Hokfelt Cocaine- and amphetamine-regulated transcript in the rat vagus nerve: A putative mediator of cholecystokinin-induced satiety PNAS, November 9, 1999; 96(23): 13506 - 13511. [Abstract] [Full Text] [PDF]

				A. M. Pilling The Role of the Toxicologic Pathologist in the Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals Toxicol Pathol, November 1, 1999; 27(6): 678 - 688. [Abstract] [PDF]

S. Schulz, H. Siem