Goat anti mouse igg biotin conjugate

The localization and distribution of PrP C in the brains of Tga20xN158D mice was analyzed by immunohistochemistry. Brains from TgN158D mice were used as controls. Serial paraffinembedded sections were incubated with a peroxidase blocking reagent (Dako) for 20 min followed by hydrated autoclaving at 100°C in citrate buffer for 30 min. Immunodetection was performed overnight at 4°C using SAF32 (1:1,000; SPI-Bio) and 5C6 (1:1,000) anti-PrP monoclonal antibodies. The anti-mouse Envision polymer (Dako) was used as the visualization system and DAB (diaminobenzidine, Dako) as the chromogen.
The localization of N158D PrP was analyzed using immunofluorescence and confocal imaging. Immunofluorescence staining was performed as described previously [26] (link), with specific modifications to adapt the protocol to paraffin-embedded samples. Paraffinembedded tissue sections from TgN158D mice were deparaffinated and rehydrated and then blocked with 1% H2O2 for 30 min. After pretreatment with 0,1% Triton X-100 for 3 h at room temperature, samples were subjected to hydrated autoclaving and incubated with SAF32 antibody (1:100) followed by a goat anti-mouse IgG biotin conjugate (1:100; Invitrogen) and an Alexa fluor 594 streptavidin conjugate (1:1000; Invitrogen). Sections were analyzed using a Zeiss laser-scanning confocal microscope LSM 510 (Carl Zeiss MicroImaging).

The histological localization and distribution of cellular PrP in the brains of both TgVole-N159D and TgVole mice was analyzed by immunohistochemistry as previously described [20] (link). Briefly, paraffin-embedded brain sections were incubated with a 1% peroxidase solution for 20 min followed by hydrated autoclaving at 100°C in citrate buffer for 30 min. PrP immunodetection was performed overnight at 4°C using SAF84
(1:1,000; Cayman Chemical) antibody. The anti-mouse Envision polymer (Dako) was used as the visualization system and DAB (diaminobenzidine, Dako) as the chromogen.
The cellular localization of PrP was further analyzed in the brains of TgVole-N159D and TgVole mice using immunofluorescence and confocal imaging. The immunofluorescence staining was performed as described previously [20] (link). Paraffinembedded tissue sections from TgVole-N159D and TgVole mice were pre-treated with 1% peroxidase for 30 min. Sections were subsequently permeabilized with 0,1% Triton X-100 for 3 h at room temperature and subjected to hydrated autoclaving.
Immunodetection was carried out with SAF84 antibody (1:1000) followed by a goat antimouse IgG biotin conjugate (1:100; Invitrogen) and an Alexa fluor 594 streptavidin conjugate (1:1000; Invitrogen). Sections were observed under a Zeiss laser-scanning confocal microscope LSM 510 (Carl Zeiss MicroImaging).