In a Leica BOND RX autostainer, deparaffinize tonsil tissue that is placed on a charged coverslip, wash (100% ethanol ×3, Leica BOND Wash solution ×3), antigen retrieval (Leica ER2 solution, EDTA pH 9.0) for 20 min at 95 °C, wash 5× with Leica BOND Wash solution, Leica 3-4% hydrogen peroxide peroxidase blocking solution (5 min), wash 3× with Leica BOND Wash solution (0 min per wash), staining with CD3e (1;200 dilution, clone, Abcam, cat. no. ab52959) primary antibody diluted in Abcam antibody diluent (Abcam, cat. no. ab64211) for 30 min, wash 3× with Leica BOND Wash solution (2 min per wash), incubate with goat anti-rabbit secondary antibody HRP conjugate ready-to-use solution (Leica, cat. no. PV6119) (8 min), wash 3× with Leica BOND Wash solution (2 min per wash), incubation with 500 nM tyramide-barcode and 500 nM tyramide-barcode in TE buffer pH 8.0 (15 min), incubation in DAPI solution (Akoya, cat. no. FP1490) diluted 3 drops per 1 mL of TBS pH 8.0 for 5 min, wash 1× with Leica BOND Wash solution (2 min per wash), collect images.
Leica bond wash solution
Manufactured by Leica camera
The Leica BOND Wash solution is a laboratory reagent designed for use with Leica's BOND automated staining system. It is a buffered aqueous solution that is used to wash and rinse slides during the staining process. The core function of the Leica BOND Wash solution is to facilitate the removal of excess reagents and maintain the appropriate pH and ionic conditions for the staining protocol.
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2 protocols using leica bond wash solution
1
Immunohistochemical Staining of Tonsil Tissue
2
Multiplex Immunohistochemistry of Tonsil Tissue
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In a Leica BOND RX autostainer, deparaffinize tonsil tissue that is placed on a charged coverslip, wash (100% ethanol ×3, Leica BOND Wash solution ×3), perform antigen retrieval (Leica ER2 solution, EDTA pH 9.0) for 20 min at 95 °C, wash 5× with Leica BOND Wash solution, incubate with Leica 3-4% hydrogen peroxide peroxidase blocking solution (5 min), wash 3× with Leica BOND Wash solution (0 min per wash), stain with CD3e (1;200 dilution, clone, Abcam, cat. no. ab52959) primary antibody diluted in Abcam antibody diluent (Abcam, cat. no. ab64211) for 30 min, wash 3× with Leica BOND Wash solution (2 min per wash), incubate with goat anti-rabbit secondary antibody HRP conjugate ready-to-use solution (Leica, cat. no. PV6119) (8 min), wash 3× with Leica BOND Wash solution (2 min per wash), incubate with 500 nM tyramide-barcode in TE buffer pH 8.0 (15 min), wash 3× with Leica BOND Wash solution (2 min per wash), incubate with 500 nM complementary barcode conjugated to Cy5 in TE buffer pH 8.0 (15 min), wash 3× with Leica BOND Wash solution (2 min per wash), incubate in DAPI solution (Akoya, cat. no. FP1490) diluted 3 drops per 1 mL of TBS pH 8.0 for 5 min, wash 1× with Leica BOND Wash solution (2 min per wash), collect images.
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