Enzyme-Labeled Antibodies: Preparation and Application for the Localization of Antigens

Pk, Nakane; Gb, Pierce

doi:10.1177/14.12.929

Cited by 934 publications

(243 citation statements)

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“…Briefly deparaffinized tissue sections obtained from 3 patients were incubated in a 10% solution of H 2 O 2 in 0.05 M Tris-HCl buffer (pH 7.6) for 5 h. Then an indirect immunostaining method (Nakane and Pierce, 1966) was applied to these specimens. An antigen retrieval method of pre-preparation of the specimen using 0.1% pronase for 5 minutes was also required for immunostaining with vWF.…”

Section: Light Microscopic Immunohistochemistrymentioning

confidence: 99%

Immunohistochemical and ultrastructural investigation of the human vestibular dark cell area: Roles of subepithelial capillaries and T lymphocyte-melanophage interaction in an immune surveillance system

et al. 1997

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Section: Light Microscopic Immunohistochemistrymentioning

confidence: 99%

Immunohistochemical and ultrastructural investigation of the human vestibular dark cell area: Roles of subepithelial capillaries and T lymphocyte-melanophage interaction in an immune surveillance system

et al. 1997

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“…After acetone fixation, sections were processedas described by Nakane and Pierce (40) . As the second antibody, swine-anti-rabbit IgG conjugated to horseradish peroxidase (Dakopatts, Denmark) was used and 3-amino-9-ethylcarbazole was used as a substrate.…”

Section: Detection Of Lung Metastasesmentioning

confidence: 99%

Metastatic behavior of human melanoma cell lines in nude mice correlates with urokinase-type plasminogen activator, its type-1 inhibitor, and urokinase-mediated matrix degradation.

Quax¹,

Muijen²,

Weening-Verhoeff³

et al. 1991

The Journal of Cell Biology

133

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Abstract. Five out of six human melanoma cell lines tested were able to degrade in vitro a smooth muscle cell extracellular matrix in a plasmin-dependent way. In three of these five cell lines, this process was mediated by tissue-type plasminogen activator (t-PA) and in the other two cell lines by urokinase-type plasminogen activator (u-PA).All melanoma cell lines produced t-PA mRNA and protein, whereas only the two cell lines showing u-PAmediated matrix degradation produced u-PA mRNA and protein . These latter cell lines also produced plasminogen activator inhibitor type-1 (PAI-1) and type-2 (PAI-2) mRNA and protein . u-PA receptor (u-PA-R) mRNA and binding of radiolabeled u-PA was found in all melanoma cell lines. The metastatic capacity of these cell lines was studied in nude mice. All cell lines were able to develop primary tumors at the sub-RING metastasis, tumor cells must penetrate basement membranes and interstitial tissues, when they detach from the primary tumor and intravasate into the circulation, and later when they extravasate at the site formation of the secondary tumor. This means that these tumor cells should express the right panel and adequate levels of proteolytic enzymes to degrade the extracellular matrix (15,16,24,34,49,57) . In addition, these enzymes may have a function in the process of angiogenesis when endothelial cells grow invasively into the newly formed tumor and form new blood vessels (25,38) . The serine protease plasmin is one ofthe major enzymes believed to be involved in such proteolytic processes (15,16,24,42,43) . Plasmin has a broad substrate specificity and can digest most of the components of the extracellular matrix including the basement membrane, either directly or by activation of proenzymes of metalloproteinases, like type IV collagenase or interstitial collagenase (23,39,57) . Plasmin is formed by a conversion of the zymogen plasminogen, which is regulated by plasminogen activators. Two distinct plasminogen activators The two u-PA and PAM producing cell lines showed the highest frequency to form spontaneous lung metastases after subcutaneous inoculation, whereas five of the six cell lines formed lung colonies after intravenous inoculation .In conclusion, u-PA mediated matrix degradation in vitro and production of u-PA and PAI-1 by human melanoma cell lines correlated with their ability to form spontaneous lung metastasis in nude mice. No correlation was found with the ability to form lung colonies after intravenous injection. These findings suggest a role for u-PA and PAI-1 in a relatively early stage of melanoma metastasis.are known, the tissue-type (t-PA),' and the urokinase-type (u-PA) . The activity ofthe activators can be regulated by interactions with specific inhibitors, of which two have been described, type 1(PAI-1) and type 2 (PAI-2) (33, 53) . In addition, u-PA and its proenzyme, pro-u-PA, can be localized at cell surfaces by binding through their growth factor domain to a specific receptor (u-PAR) (2,8,12,41,51,55,56,62) .To study the role of plasminoge...

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“…An alternative to fluoresceinated stains is the enzyme-based immunohistochemical methods. 21 Immunoenzyme stains are permanent preparations visible with ordinary light microscopy and may be simultaneously counterstained for optimal demonstration of the histologic lesions, which may assist in establishing the pathogenicity of the agent in the case. 15 This advantage may be particularly important in assessing the contribution of the PI3 virus, given the uncertainty of this virus in BRD cases.…”

mentioning

confidence: 99%

Monoclonal and Polyclonal Antibodies for Immunohistochemical Detection of Bovine Parainfluenza Type 3 Virus in Frozen and Formalin-Fixed Paraffin-Embedded Tissues

et al. 1992

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Abstract. Accurate identification of bovine Parainfluenza type 3 virus in bovine respiratory disease requires dependable, sensitive, and specific techniques for detection in affected animals. Immunohistochemical testing can be a rapid and reliable means of demonstration of virus in tissues from suspect cases; however, this procedure is dependent upon the quality of the antisera directed against the viral antigens. The production of rabbit polyclonal and murine monoclonal antibodies directed against bovine Parainfluenza type 3 virus and techniques for their use in fresh-frozen and formalin-fixed paraffin-embedded tissues in immunofluorescence and immunoperoxidase-based immunohistochemical tests are described.

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Enzyme-Labeled Antibodies: Preparation and Application for the Localization of Antigens

Cited by 934 publications

References 0 publications

Immunohistochemical and ultrastructural investigation of the human vestibular dark cell area: Roles of subepithelial capillaries and T lymphocyte-melanophage interaction in an immune surveillance system

Immunohistochemical and ultrastructural investigation of the human vestibular dark cell area: Roles of subepithelial capillaries and T lymphocyte-melanophage interaction in an immune surveillance system

Metastatic behavior of human melanoma cell lines in nude mice correlates with urokinase-type plasminogen activator, its type-1 inhibitor, and urokinase-mediated matrix degradation.

Monoclonal and Polyclonal Antibodies for Immunohistochemical Detection of Bovine Parainfluenza Type 3 Virus in Frozen and Formalin-Fixed Paraffin-Embedded Tissues

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