scholarly journals Immunohistochemical detection of the androgen receptor with monoclonal antibody F39.4 in routinely processed, paraffin-embedded human tissues after microwave pre-treatment.

1994 ◽  
Vol 42 (8) ◽  
pp. 1169-1175 ◽  
Author(s):  
P J Janssen ◽  
A O Brinkmann ◽  
W J Boersma ◽  
T H Van der Kwast
Keyword(s):  
Monoclonal Antibody ◽  
Androgen Receptor ◽  
Staining Intensity ◽  
Antigen Retrieval ◽  
Nuclear Staining ◽  
Immunohistochemical Detection ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Long Term Storage ◽  
Pre Treatment

We describe the immunohistochemical detection of the human androgen receptor (AR) in routinely processed, paraffin-embedded tissue with the monoclonal antibody (MAb) F39.4. Deparaffinized sections were heated in a microwave oven for antigen retrieval. A panel of human male- and female-derived tissues was investigated. We observed a nuclear staining pattern consistent with previous results on frozen sections. Moreover, we studied the possibility of detecting AR in prolonged formalin-fixed tissue and in paraffin-embedded archival material. After prolonged fixation times or long-term storage of paraffin-embedded tissue, the staining intensity for the AR did not deteriorate. Blocking experiments with the specific synthetic peptides demonstrated the specificity of this technique. We conclude that this method is specific, allows retrospective AR studies, and offers optimally preserved morphology.

scholarly journals Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections.

1991 ◽  
Vol 39 (6) ◽  
pp. 741-748 ◽  
Author(s):  
S R Shi ◽  
M E Key ◽  
K L Kalra
Keyword(s):  
Polyclonal Antibodies ◽  
Antigen Retrieval ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Pre Treatment ◽  
Immunohistochemical Techniques ◽  
Formalin Fixed

We describe a new approach for retrieval of antigens from formalin-fixed, paraffin-embedded tissues and their subsequent staining by immunohistochemical techniques. This method of antigen retrieval is based on microwave heating of tissue sections attached to microscope slides to temperatures up to 100 degrees C in the presence of metal solutions. Among 52 monoclonal and polyclonal antibodies tested by this method, 39 antibodies demonstrated a significant increase in immunostaining, nine antibodies showed no change, and four antibodies showed reduced immunostaining. In particular, excellent immunostaining results were obtained with a monoclonal antibody to vimentin as well as several different keratin antibodies on routine formalin-fixed tissue sections after pre-treatment of the slides with this method. These results showed that after antigen retrieval: (a) enzyme predigestion of tissues could be omitted; (b) incubation times of primary antibodies could be significantly reduced, or dilutions of primary antibodies could be increased; (c) adequate staining could be achieved in long-term formalin-fixed tissues that failed to stain by conventional methods; and (d) certain antibodies which were typically unreactive with formalin-fixed tissues gave excellent staining.

Effects of antigen retrieval by microwave heating in formalin-fixed tissue sections on a broad panel of antibodies

1994 ◽  
Vol 102 (3) ◽  
pp. 165-172 ◽  
Author(s):  
R. von Wasielewski ◽  
M. Werner ◽  
M. Nolte ◽  
L. Wilkens ◽  
A. Georgii
Keyword(s):  
Microwave Heating ◽  
Antigen Retrieval ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Tissue Sections ◽  
Formalin Fixed

scholarly journals Protease Antigen Recovery Decreases the Specificity of Bromodeoxyuridine Detection In Formalin-fixed Tissue

1997 ◽  
Vol 45 (8) ◽  
pp. 1165-1170 ◽  
Author(s):  
Philip M. Bak ◽  
Ralph J. Panos
Keyword(s):  
Cellular Proliferation ◽  
Enzymatic Digestion ◽  
Antigen Retrieval ◽  
Alveolar Type ◽  
Positive Staining ◽  
Specific Staining ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Protease Digestion ◽  
Formalin Fixed

Incorporation of halogenated nucleotide analogues is often used to assess DNA synthesis and to quantitate cellular proliferation. Multiple antibodies have been developed to bromodeoxyuridine (BrdUrd) and it is the most frequently utilized substrate. Because the immunodetection of incorporated BrdUrd requires DNA denaturation or nuclease digestion, most of these antibodies are not reactive in tissues or cells fixed with crosslinking agents. Antigen retrieval techniques utilizing protease digestion restore BrdUrd antigenicity and permit the detection of BrdUrd in formalin-fixed tissue. However, during the development of a double label immunohistochemical protocol to quantitate proliferating alveolar Type II cells, we noted nucleus-specific staining in lung sections from animals that had not received BrdUrd. Therefore, we systematically analyzed the specificity of the immunohistochemical detection of incorporated BrdUrd in formalin-fixed tissue after protease digestion. Enzymatic antigen recovery diminished the specificity of the BrdUrd reaction product and caused false-positive staining with the BU-1, B44, and BR3 monoclonal antibodies. Staining was less prominent with Bu20a but was more specific. Protease antigen recovery may decrease the specificity of BrdUrd immunodetection. Appropriate controls are required when enzymatic digestion is used to detect incorporated BrdUrd in formalin-fixed tissue. The type and duration of fixation, antibody to BrdUrd, protease, and tissue may affect the specificity of the staining pattern. (J Histochem Cytochem 45:1165–1170, 1997)

Monoclonal Antibody-based Immunohistochemical Detection of Bovine Viral Diarrhea Virus in Formalin-fixed, Paraffin-embedded Tissues

1992 ◽  
Vol 29 (1) ◽  
pp. 27-32 ◽  
Author(s):  
D. M. Haines ◽  
E. G. Clark ◽  
E. J. Dubovi
Keyword(s):  
Monoclonal Antibody ◽  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Immunohistochemical Detection ◽  
Fixed Tissue ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Mucosal Disease ◽  
Viral Diarrhea Virus ◽  
Formalin Fixed

Thirty-two monoclonal antibodies directed against epitopes on bovine viral diarrhea virus proteins and glycoproteins were tested for immunohistochemical reactivity with bovine viral diarrhea virus in formalin-fixed and paraffin-embedded tissues from 45 cases of bovine viral diarrhea virus-associated mucosal disease. Only one antibody, designated 15C5, which reacts with the 48-kD glycoprotein of bovine viral diarrhea virus, detected an epitope preserved in these specimens. Monoclonal antibody 15C5 and a polyclonal antibody to bovine viral diarrhea virus successfully detected bovine viral diarrhea viral antigens in 44/45 cases of mucosal disease and did not react with formalin-fixed tissues from 30 uninfected cattle. Monoclonal antibody-based immunohistochemical detection of bovine viral diarrhea virus in routinely fixed tissue specimens has advantages over other currently available techniques in terms of the convenience of specimen submission, the relative ease of method standardization, and the rapidity of the test, and by enabling identification of the virus in association with specific tissues, cell types, and histologic lesions.

Standardization of Immunohistochemistry Based on Antigen Retrieval Technique for Routine Formalin-fixed Tissue Sections

1998 ◽  
Vol 6 (2) ◽  
pp. 89-96 ◽  
Author(s):  
Shan-Rong Shi ◽  
Richard J. Cote ◽  
Benjaporn Chaiwun ◽  
Lillian L. Young ◽  
Yan Shi ◽  
...  
Keyword(s):  
Antigen Retrieval ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Tissue Sections ◽  
Retrieval Technique ◽  
Formalin Fixed
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