Autoclave Antigen Retrieval Technique for Immunohistochemical Staining of Androgen Receptor in Formalin-Fixed Paraffin Sections of Human Prostate.

Ehara, Hidetoshi; Deguchi, Takashi; Koji, Takehiko; Yang, Muzheng; S, Itó; Kawada, Yukimichi; Nakane, Paul K.

doi:10.1267/ahc.29.311

Cited by 16 publications

(13 citation statements)

References 18 publications

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“…Deparaffinized and rehydrated sections were immersed in 0.01 M citrate buffer (pH 6.0) and then autoclaved at 120 °C for 10 min as described previously [20]. The sections were then immersed in 0.3% (v/v) H2O2 in methanol to eliminate endogenous peroxidase activity and were preincubated with 500 jig/ml goat IgG and 1 % bovine serum albumin (BSA) in PBS for 1 h at room temperature to block nonspecific binding of antibodies.…”

Section: Introductionmentioning

confidence: 99%

Evidence for an Inverse Relationship between Apoptosis and Inducible Nitric Oxide Synthase Expression in Rat Granulosa Cells: A Possible Role of Nitric Oxide in Ovarian Follicle Atresia.

et al. 1998

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Section: Introductionmentioning

confidence: 99%

Evidence for an Inverse Relationship between Apoptosis and Inducible Nitric Oxide Synthase Expression in Rat Granulosa Cells: A Possible Role of Nitric Oxide in Ovarian Follicle Atresia.

et al. 1998

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“…Thus, the molarity of the sodium citrate buffer 39 was stable, reducing background effects. In other studies it was reported that an AR solution with 0.01M sodium citrate at pH 6.0 showed satisfactory results, preserving cell morphology when compared to other buffers with high pH, or containing EDTA 13,36,40,41 .…”

Section: Discussionmentioning

confidence: 90%

Comparative analysis of two immunohistochemical methods for antigen retrieval in the optical lobe of the honeybee Apis mellifera: Myosin-v assay

et al. 2010

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The present study compared two heating methods currently used for antigen retrieval (AR) immunostaining: the microwave oven and the steam cooker. Myosin-V, a molecular motor involved in vesicle transport, was used as a neuronal marker in honeybee Apis mellifera brains fixed in formalin. Overall, the steam cooker showed the most satisfactory AR results. At 100 o C, tissue morphology was maintained and revealed epitope recovery, while evaporation of the AR solution was markedly reduced; this is important for stabilizing the sodium citrate molarity of the AR buffer and reducing background effects. Standardization of heat-mediated AR of formalin-fixed and paraffin-embedded tissue sections results in more reliable immunostaining of the honeybee brain.

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“…Enzymatic treatment causes the disruption of the crosslinks between the epitopes and the fixative molecules, which have modified the structures of the protein [33]. The enzymatic treatment immuno-reactivity is further enhanced by presence of number of lysine, arginine or asparagine amino acid residues [33], and since laminin is rich in such amino acid residues [34] Ehara et al [37] suggested that for most Antigens, HIER with sodium citrate buffer will give satisfactory results and good cell morphology when compared with buffers with higher pH such as Tris-EDTA. Kim et al [38] stated that the basic buffer solution like Tris-EDTA seems to be more efficient in retrieving antigens than citrate buffer.…”

Section: Discussionmentioning

confidence: 99%

A Combination of Antigen Retrieval Methods Significantly Enhances the Unmasking of Tedious Antigens of Lymphangiogenesis and Angiogenesis in Canine Mammary Tumour

Singh¹,

Sood²

2017

JBM

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The use of immunohistochemical techniques has enhanced the diagnostic intensities in pathology. The sensitivity and specificity of any immunohistochemical protocol mainly depends on the ability to unmask antigen from formalin fixed tissue. Various combinations of antigen retrieval protocols have been used to unmask antigen efficiently but none has been established as a flawless one. These protocols either have one or the other lacunae, lacking repeatability under same conditions. In our study, we tried to develop a standardized immunohistochemical protocol for the identification of blood and lymphatic vessels in tissue sections of canine mammary tumour (CMT) using recently identified markers. The combined effects of antigen retrieval (AR) methods including pH shock by Heat-induced epitope retrieval (HIER) and Protease-induced epitope retrieval (PIER) were found highly effective in retrieval of tedious antigens of lymphangiogenesis and angiogenesis. The use of combined antigen retrieval technique for the unmasking of antigens from over fixed, old formalin fixed paraffin embedded (FFPE) canine mammary tumour sections resulted in efficient unmasking of epitopes when compared with the individual antigen retrieval methods.

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Autoclave Antigen Retrieval Technique for Immunohistochemical Staining of Androgen Receptor in Formalin-Fixed Paraffin Sections of Human Prostate.

Cited by 16 publications

References 18 publications

Evidence for an Inverse Relationship between Apoptosis and Inducible Nitric Oxide Synthase Expression in Rat Granulosa Cells: A Possible Role of Nitric Oxide in Ovarian Follicle Atresia.

Evidence for an Inverse Relationship between Apoptosis and Inducible Nitric Oxide Synthase Expression in Rat Granulosa Cells: A Possible Role of Nitric Oxide in Ovarian Follicle Atresia.

Comparative analysis of two immunohistochemical methods for antigen retrieval in the optical lobe of the honeybee Apis mellifera: Myosin-v assay

A Combination of Antigen Retrieval Methods Significantly Enhances the Unmasking of Tedious Antigens of Lymphangiogenesis and Angiogenesis in Canine Mammary Tumour

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