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Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures

SM Hsu, L Raine and H Fanger

The use of avidin-biotin interaction in immunoenzymatic techniques provides a simple and sensitive method to localize antigens in formalin- fixed tissues. Among the several staining procedures available, the ABC method, which involves an application of biotin-labeled secondary antibody followed by the addition of avidin-biotin-peroxidase complex, gives a superior result when compared to the unlabeled antibody method. The availability of biotin-binding sites in the complex is created by the incubation of a relative excess of avidin with biotin-labeled peroxidase. During formation of the complex, avidin acts as a bridge between biotin-labeled peroxidase molecules; and biotin-labeled peroxidase molecules, which contains several biotin moieties, serve as a link between the avidin molecules. Consequently, a "lattice" complex containing several peroxidase molecules is likely formed. Binding of this complex to the biotin moieties associated with secondary antibody results in a high staining intensity.

Volume 29, Issue 4, pp. 577-580, 04/01/1981
Copyright © 1981 by The Histochemical Society


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J. Histochem. Cytochem., April 1, 2007; 55(4): 375 - 386.
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F. Groenman, M. Rutter, I. Caniggia, D. Tibboel, and M. Post
Hypoxia-inducible Factors in the First Trimester Human Lung
J. Histochem. Cytochem., April 1, 2007; 55(4): 355 - 363.
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T.-K. Lee, J. H. Lois, J. H. Troupe, T. D. Wilson, and B. J. Yates
Transneuronal tracing of neural pathways that regulate hindlimb muscle blood flow
Am J Physiol Regulatory Integrative Comp Physiol, April 1, 2007; 292(4): R1532 - R1541.
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 BloodHome page
K. J. Aichberger, M. Mayerhofer, K. V. Gleixner, M.-T. Krauth, A. Gruze, W. F. Pickl, V. Wacheck, E. Selzer, L. Mullauer, H. Agis, et al.
Identification of MCL1 as a novel target in neoplastic mast cells in systemic mastocytosis: inhibition of mast cell survival by MCL1 antisense oligonucleotides and synergism with PKC412
Blood, April 1, 2007; 109(7): 3031 - 3041.
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N. Cuesta, Q. M. Nhu, E. Zudaire, S. Polumuri, F. Cuttitta, and S. N. Vogel
IFN Regulatory Factor-2 Regulates Macrophage Apoptosis through a STAT1/3- and Caspase-1-Dependent Mechanism
J. Immunol., March 15, 2007; 178(6): 3602 - 3611.
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 BloodHome page
N. Harir, C. Pecquet, M. Kerenyi, K. Sonneck, B. Kovacic, R. Nyga, M. Brevet, I. Dhennin, V. Gouilleux-Gruart, H. Beug, et al.
Constitutive activation of Stat5 promotes its cytoplasmic localization and association with PI3-kinase in myeloid leukemias
Blood, February 15, 2007; 109(4): 1678 - 1686.
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K. S. Smith and K. C. Berridge
Opioid Limbic Circuit for Reward: Interaction between Hedonic Hotspots of Nucleus Accumbens and Ventral Pallidum
J. Neurosci., February 14, 2007; 27(7): 1594 - 1605.
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 J DAIRY SCIHome page
A. Rabot, F. Sinowatz, B. Berisha, H. H. D. Meyer, and D. Schams
Expression and Localization of Extracellular Matrix-Degrading Proteinases and Their Inhibitors in the Bovine Mammary Gland During Development, Function, and Involution
J Dairy Sci, February 1, 2007; 90(2): 740 - 748.
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 Arch DermatolHome page
G. Maire, S. Fraitag, L. Galmiche, F. Keslair, N. Ebran, M.-J. Terrier-Lacombe, Y. de Prost, and F. Pedeutour
A Clinical, Histologic, and Molecular Study of 9 Cases of Congenital Dermatofibrosarcoma Protuberans
Arch Dermatol, February 1, 2007; 143(2): 203 - 210.
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 EndocrinologyHome page
M.-L. Baudet, B. Martin, Z. Hassanali, E. Parker, E. J. Sanders, and S. Harvey
Expression, Translation, and Localization of a Novel, Small Growth Hormone Variant
Endocrinology, January 1, 2007; 148(1): 103 - 115.
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 Cereb CortexHome page
S. M. Eggan and D. A. Lewis
Immunocytochemical Distribution of the Cannabinoid CB1 Receptor in the Primate Neocortex: A Regional and Laminar Analysis
Cereb Cortex, January 1, 2007; 17(1): 175 - 191.
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 BrainHome page
F. Loup, F. Picard, V. M. Andre, P. Kehrli, Y. Yonekawa, H.-G. Wieser, and J.-M. Fritschy
Altered expression of {alpha}3-containing GABAA receptors in the neocortex of patients with focal epilepsy
Brain, December 1, 2006; 129(12): 3277 - 3289.
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 BloodHome page
K. J. Aichberger, M. Mayerhofer, A. Vales, M.-T. Krauth, K. V. Gleixner, M. Bilban, H. Esterbauer, K. Sonneck, S. Florian, S. Derdak, et al.
The CML-related oncoprotein BCR/ABL induces expression of histidine decarboxylase (HDC) and the synthesis of histamine in leukemic cells
Blood, November 15, 2006; 108(10): 3538 - 3547.
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 BrainHome page
E. Capetillo-Zarate, M. Staufenbiel, D. Abramowski, C. Haass, A. Escher, C. Stadelmann, H. Yamaguchi, O. D. Wiestler, and D. R. Thal
Selective vulnerability of different types of commissural neurons for amyloid {beta}-protein-induced neurodegeneration in APP23 mice correlates with dendritic tree morphology
Brain, November 1, 2006; 129(11): 2992 - 3005.
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 Clin. Cancer Res.Home page
Y. Tajima, K. Yamazaki, R. Makino, N. Nishino, S. Aoki, M. Kato, K. Morohara, T. Kaetsu, and M. Kusano
Gastric and intestinal phenotypic marker expression in early differentiated-type tumors of the stomach: clinicopathologic significance and genetic background.
Clin. Cancer Res., November 1, 2006; 12(21): 6469 - 6479.
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 J EndocrinolHome page
A. Chamson-Reig, S. M Thyssen, E. Arany, and D. J Hill
Altered pancreatic morphology in the offspring of pregnant rats given reduced dietary protein is time and gender specific.
J. Endocrinol., October 1, 2006; 191(1): 83 - 92.
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 FASEB J.Home page
T. Yakovleva, I. Bazov, G. Cebers, Z. Marinova, Y. Hara, A. Ahmed, M. Vlaskovska, B. Johansson, U. Hochgeschwender, I. N. Singh, et al.
Prodynorphin storage and processing in axon terminals and dendrites
FASEB J, October 1, 2006; 20(12): 2124 - 2126.
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 Toxicol SciHome page
M. D. Boudreau, H. W. Taylor, D. G. Baker, and J. C. Means
Dietary Exposure to 2-Aminoanthracene Induces Morphological and Immunocytochemical Changes in Pancreatic Tissues of Fisher-344 Rats
Toxicol. Sci., September 1, 2006; 93(1): 50 - 61.
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 J. Clin. Pathol.Home page
M Nowicki, D Ostalska-Nowicka, A Konwerska, and B Miskowiak
The predicting role of substance P in the neoplastic transformation of the hypoplastic bone marrow.
J. Clin. Pathol., September 1, 2006; 59(9): 935 - 941.
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 J. Clin. Pathol.Home page
D Ostalska-Nowicka, J Zachwieja, M Nowicki, E Kaczmarek, A Siwinska, and M Witt
Ezrin--a useful factor in the prognosis of nephrotic syndrome in children: an immunohistochemical approach
J. Clin. Pathol., September 1, 2006; 59(9): 916 - 920.
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 Clin. Cancer Res.Home page
H.-J. Heidebrecht, A. Claviez, M. L. Kruse, M. Pollmann, F. Buck, S. Harder, M. Tiemann, W. Dorffel, and R. Parwaresch
Characterization and Expression of CT45 in Hodgkin's Lymphoma.
Clin. Cancer Res., August 15, 2006; 12(16): 4804 - 4811.
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 Mol. Cell. Biol.Home page
I. Soeiro, A. Mohamedali, H. M. Romanska, N. C. Lea, E. S. Child, J. Glassford, S. J. Orr, C. Roberts, K. N. Naresh, E.-N. Lalani, et al.
p27Kip1 and p130 Cooperate To Regulate Hematopoietic Cell Proliferation In Vivo.
Mol. Cell. Biol., August 1, 2006; 26(16): 6170 - 6184.
[Abstract] [Full Text] [PDF]

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 JCOHome page
I. F. Pollack, R. L. Hamilton, R. W. Sobol, J. Burnham, A. J. Yates, E. J. Holmes, T. Zhou, and J. L. Finlay
O6-Methylguanine-DNA Methyltransferase Expression Strongly Correlates With Outcome in Childhood Malignant Gliomas: Results From the CCG-945 Cohort
J. Clin. Oncol., July 20, 2006; 24(21): 3431 - 3437.
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 Proc. Natl. Acad. Sci. USAHome page
M. B. Cachon-Gonzalez, S. Z. Wang, A. Lynch, R. Ziegler, S. H. Cheng, and T. M. Cox
Effective gene therapy in an authentic model of Tay-Sachs-related diseases
PNAS, July 5, 2006; 103(27): 10373 - 10378.
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 INT J SURG PATHOLHome page
L. Posligua, F. Anatelli, L. P. Dehner, and J. D. Pfeifer
Primary Peritoneal Epithelioid Hemangioendothelioma
International Journal of Surgical Pathology, July 1, 2006; 14(3): 257 - 267.
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R. Klopfleisch, B. G. Klupp, W. Fuchs, M. Kopp, J. P. Teifke, and T. C. Mettenleiter
Influence of pseudorabies virus proteins on neuroinvasion and neurovirulence in mice.
J. Virol., June 1, 2006; 80(11): 5571 - 5576.
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 Mol. Cell. ProteomicsHome page
S. Ek, U. Andreasson, S. Hober, C. Kampf, F. Ponten, M. Uhlen, H. Merz, and C. A. K. Borrebaeck
From Gene Expression Analysis to Tissue Microarrays: A Rational Approach to Identify Therapeutic and Diagnostic Targets in Lymphoid Malignancies
Mol. Cell. Proteomics, June 1, 2006; 5(6): 1072 - 1081.
[Abstract] [Full Text] [PDF]

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 CarcinogenesisHome page
T. P. Ong, R. Heidor, A. de Conti, M. L. Z. Dagli, and F. S. Moreno
Farnesol and geraniol chemopreventive activities during the initial phases of hepatocarcinogenesis involve similar actions on cell proliferation and DNA damage, but distinct actions on apoptosis, plasma cholesterol and HMGCoA reductase
Carcinogenesis, June 1, 2006; 27(6): 1194 - 1203.
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 Clin. Cancer Res.Home page
D. Furlan, I. Carnevali, B. Marcomini, R. Cerutti, E. Dainese, C. Capella, and C. Riva
The High Frequency of De novo Promoter Methylation in Synchronous Primary Endometrial and Ovarian Carcinomas.
Clin. Cancer Res., June 1, 2006; 12(11): 3329 - 3336.
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 EndocrinologyHome page
S. Thyssen, E. Arany, and D. J. Hill
Ontogeny of Regeneration of {beta}-Cells in the Neonatal Rat after Treatment with Streptozotocin
Endocrinology, May 1, 2006; 147(5): 2346 - 2356.
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 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
M. van Tuyl, J. Liu, F. Groenman, R. Ridsdale, R. N. N. Han, V. Venkatesh, D. Tibboel, and M. Post
Iroquois genes influence proximo-distal morphogenesis during rat lung development
Am J Physiol Lung Cell Mol Physiol, April 1, 2006; 290(4): L777 - L789.
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H Agis, M-T Krauth, I Mosberger, L Mullauer, I Simonitsch-Klupp, L B Schwartz, D Printz, A Bohm, G Fritsch, H-P Horny, et al.
Enumeration and immunohistochemical characterisation of bone marrow basophils in myeloproliferative disorders using the basophil specific monoclonal antibody 2D7
J. Clin. Pathol., April 1, 2006; 59(4): 396 - 402.
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 J. Clin. Pathol.Home page
H-P Horny, K Sotlar, F Stellmacher, P Valent, and J Grabbe
An unusual case of systemic mastocytosis associated with chronic lymphocytic leukaemia (SM-CLL).
J. Clin. Pathol., March 1, 2006; 59(3): 264 - 268.
[Abstract] [Full Text] [PDF]

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 J. Clin. Pathol.Home page
H-P Horny, K Sotlar, F Stellmacher, M Krokowski, H Agis, L B Schwartz, and P Valent
The tryptase positive compact round cell infiltrate of the bone marrow (TROCI-BM): a novel histopathological finding requiring the application of lineage specific markers.
J. Clin. Pathol., March 1, 2006; 59(3): 298 - 302.
[Abstract] [Full Text] [PDF]

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 BrainHome page
M. V. Solbrig, R. Adrian, J. Baratta, J. C. Lauterborn, and G. F. Koob
Kappa opioid control of seizures produced by a virus in an animal model
Brain, March 1, 2006; 129(3): 642 - 654.
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