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| Furuoka, Hidefumi; Yabuzoe, Atushi; Horiuchi, Motohiro; Tagawa, Yuichi; Yokoyama, Takashi; Yamakawa, Yoshio; Shinagawa, Morikazu; Sata, Tetsutaro. |
| For immunohistochemistry of the prion diseases, several pretreatment methods to enhance the immunoreactivity of human and animal abnormal proteinase-resistant prion protein (PrPSc) on the tissue sections have been employed. The method of 121 degrees C hydrated autoclaving pretreatment or the combination method of 121 degrees C hydrated autoclaving with a certain chemical reagent ( formic acid or proteinase K, etc) are now widely used. We found that an improved hydrated autoclaving method at 135 degrees C, more effectively enhanced PrPSc immunoreactivity for the antibodies recognizing the linear epitope. In addition, this method was more effective for the long-term fixation samples as compared with other previous methods. However, this modified method could... |
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Palavras-chave: Prion protein; Immunohistochemistry; Antigen retrieval; Autoclaving; Monoclonal antibody; CREUTZFELDT-JAKOB-DISEASE; MONOCLONAL-ANTIBODIES; IMMUNOCYTOCHEMISTRY; IMMUNOREACTIVITY; PROTOCOLS; FIXATION; DEPOSITS; PLAQUES; BRAIN; PRP. |
| Ano: 2005 |
URL: http://ir.obihiro.ac.jp/dspace/handle/10322/712 |
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| CALÁBRIA,LUCIANA KAREN; TEIXEIRA,RENATA ROLAND; COELHO GONÇALVES,SYBELLI MAGDA; PASSOS LIMA,ANDREIA BARCELOS; SANTOS,ANA ALICE DINIZ DOS; MARTINS,ANTÔNIO ROBERTO; ESPINDOLA,FOUED SALMEN. |
| 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 ºC, 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. |
| Tipo: Journal article |
Palavras-chave: Antigen retrieval; Microwave oven; Steam cooker. |
| Ano: 2010 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602010000100002 |
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