Intended use: For in-vitro Diagnostic Use. The BrightVision Ultimate one step component detection system peroxidase Goat Anti-Mouse/Rabbit IgG HRP and DAB, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
Summary and explanation: The BrightVision Ultimate detection system, peroxidase Goat Anti-Mouse/Rabbit HRP plus DAB, 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 Ultimate detection system detects Mouse or Rabbit 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 Ultimate plus, one detection system, Goat Anti-Mouse/Rabbit HRP (Ready-to-Use) plus DAB1. one step detection, Goat Anti-Mouse/Rabbit HRP (Ready-to-Use): 55 ml 2. DAB Solution A: Buffered H2O2 (Ready-to-Use): 83 ml4. DAB Solution B: Concentrated DAB solution: 4 ml
Storage and handling: 2-8°C and in the dark
Principle of method: BrightVision Ultimate, one step detection system, Goat Anti-Mouse/Rabbit HRP (Ready-to-Use) plus DAB.
Reagent preparation: Preparation DAB: Add 40μl DAB Solution B (one drop) to 1 ml substrate Solution A, mix well. Volume and the quality of the Bright DAB has been formulized so they also can be used in automatic stainers, when a higher volume is required.
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 or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9 Detection system, polymer Mouse/Rabbit HRP, (Labeled polymer; 30 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Substrate (DAB; 8 min.; Preparation DAB: Add 40μl DAB Solution B (one drop) to 1 ml substrate Solution A, mix well. Volume and the quality of the Bright DAB has been formulized so they also can be used in automatic stainers, when a higher volume is required.), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain, dyhydrate and coverslip (Auxiliary.
References: Shan-Rong Shi et al. Applied Immunohistochemistry & Molecular Morphology, vol 7,201-208,2003
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