Improved Immunoperoxidase Stain on Frozen Sections: An Avidin-Biotin-Peroxidase Complex (ABC) Technique

Marty, Joe; Kjeldsberg, Carl R.; Groo, Steve

doi:10.1179/his.1982.5.2.61

Cited by 12 publications

(2 citation statements)

References 5 publications

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“…Dehydrating and delipidating in 70%, 80%, 95%, and absolute ethanols (eight steps, 1 h each) were followed by clearing in either chloroform or xylene (three steps, I h each), impregnating in paraffin (Paraplast Plus, 58-60°C; three steps, I h each), and embedding in paraffin. Sections were cut at 6 pm, deparaffinized, hydrated in PBST, and stained with one or another of the MAbs for 2 h at room temperature followed by reactions for 1 h at room temperature with rabbit anti-mouse IgG or IgM or anti-rat IgG antisera (diluted 1 : IOO), biotinylated goat anti-rabbit IgG (diluted 1 : loo), and avidin-biotin-peroxidase complex (ABC) reagent (Vector Laboratories, Burlingame, CA) according to the method of Marty et al (1982).…”

Section: Immunohistologymentioning

confidence: 99%

Sites in Myelin Basic Protein that React with Monoclonal Antibodies

Hruby

Alvord

Martenson

et al. 1985

Journal of Neurochemistry

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The epitopes (antigenic sites) for seven monoclonal antibodies (MAbs) evoked in rats or mice by guinea pig or monkey myelin basic protein (BP) have been located in four different sequences of the BPs extracted from various species. Six of the MAbs were evoked by guinea pig BP. (1) One epitope, possibly a pair, is included within residues 1-14 of all BPs tested and reacts with two rat IgG MAbs. (2) A definite pair of overlapping epitopes includes the central Phe91-Phe92 sequence. One epitope is contained entirely within sequence 90-99 and reacts with a rat IgG MAb. The substitution of Ser in chicken BP for Thr97 destroys this epitope. The other epitope appears to include residues on the amino side of Phe44 and even of His32 and suggests some tertiary structure in BP. This epitope reacts with a mouse IgM MAb that does not recognize the chicken substitution. (3) The third epitope lies within residues 114-121, specifically including Trp118, and reacts with a rat IgG MAb. A cross-reacting epitope probably includes residues 44-45 in certain species (guinea pig and bovine but not rabbit). (4) Another pair of epitopes is located within residues 131-140 but is severely species-restricted. This region in guinea pig BP evoked a species-specific mouse IgM MAb. The same region in monkey BP evoked the seventh MAb, a mouse IgG, which reacts with human, chimpanzee, monkey, bovine, and rat-18.5 kDa BPs and to a lesser extent rabbit BP but not with guinea pig, pig, or chicken BPs. Some tertiary structure in guinea pig BP is also suggested by the reactivities with the IgM MAb. All of the MAbs react with myelin in histologic preparations, but the optimum method of preparation of the tissue varies with each.

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Section: Immunohistologymentioning

confidence: 99%

Sites in Myelin Basic Protein that React with Monoclonal Antibodies

Hruby

Alvord

Martenson

et al. 1985

Journal of Neurochemistry

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“…), and anti-T11 (Ortho, Raritan, NJ). Similar studies were performed on frozen tissue sections (cases 1, 2 and 3) and on tumor imprints and cell suspensions (case 1) using the ABC immunoperoxidase method (Marty et al, 1982). The same technique was used on paraffin sections from case 4 to detect intracytoplasmic immunoglobulin (CIg).…”

Section: Immunohistochemical Studiesmentioning

confidence: 99%

Multilobated B cell lymphomas. A study of 7 cases

Weiss

Kjeldsberg

Colby

et al. 1985

Hematological Oncology

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Simultaneous inhibition of endogenous avidin-binding activity and peroxidase applicable for the avidin-biotin system using monoclonal antibodies

Matsumoto

1985

Histochemistry

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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 (EPA) also makes the estimation of specific staining difficult. In the present study, several methods for the inhibition of EABA and/or EPA were examined using the avidin-biotin technique and monoclonal antibodies against murine Mac-1 and Ia antigen. Of these, the overnight incubation of sections in 40% methanol in phosphate-buffered saline containing 0.3% hydrogen peroxide gave the best result, as it inhibited EABA and EPA simultaneously without denaturating of the antigenic determinants recognized by the monoclonal antibodies.

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Improved Immunoperoxidase Stain on Frozen Sections: An Avidin-Biotin-Peroxidase Complex (ABC) Technique

Cited by 12 publications

References 5 publications

Sites in Myelin Basic Protein that React with Monoclonal Antibodies

Sites in Myelin Basic Protein that React with Monoclonal Antibodies

Multilobated B cell lymphomas. A study of 7 cases

Simultaneous inhibition of endogenous avidin-binding activity and peroxidase applicable for the avidin-biotin system using monoclonal antibodies

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