Intended use: For in-vitro Diagnostic Use. The BrightVision one step detection system Goat Anti-Rabbit IgG HRP and Goat anti-Mouse IgG AP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
Summary and explanation: The BrightVision detection system, peroxidase Goat Anti-Rabbit HRP and Goat anti-Mouse AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU.Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Rabbit and Mouse bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen.The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patient’s clinical history and/or other diagnostic tests.
Reagents provided: BrightVision, One step detection system Goat anti-Rabbit HRP / Goat anti-Mouse AP (Ready-to-use; 55 ml)
Storage and handling: 2-8°C and in the dark
Principle of method: BrightVision, one step detection system Goat Anti- Rabbit HRP and Mouse AP (Ready-to-Use).
Procedure: 1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse and/or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9 Detection system, polymer Rabbit HRP/Mouse AP, (Labeled polymer; 30 min), 10. Wash buffer (TBS buffer; 2x 5 min), 11. Substrate (DAB; see applicble IFU), 12. Wash aqua dest (Wash; 2x 2 min), 13. Substrate (Fast Red / New Fuchsin; see applicable IFU), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain and coverslip with aqueous mounting medium.
References: Shan-Rong Shi et al. Applied Immunohistochemistry & Molecular Morphology, vol 7,201-208,2005