I. Required material
Reagents
- VECTABOND™ Reagent
Buffers
II. Experimentation length
- 30 mn of sections incubation in primary antibody
- 10 mn of sections incubation in secondary antibody
- 1h10 incubation with different the reagents
- 26 mn of rinsing steps
- TOTAL :
1. Slide Preparation:
Place clean glass slides in metal slide racks. Use either good quality pre-cleaned slides or wash slides thoroughly in detergent and rinse in water. Immerse slides in acetone for 5 minutes.
2. Slide Treatment:
Prepare VECTABOND™ Reagent treatment solution by adding the entire contents (7 ml) of the bottle to 350 ml of acetone and stirring well. Remove slides from acetone, tapping rack several times to drain and immediately place in VECTABOND™ Reagent solution for 5 minutes. Remove slides and drain. Eliminate excess reagent by gently dipping them several times over 30 seconds
in deionized or distilled water (do not create bubbles), changing the water after every 5 racks. Remove rack of slides for drying. Gentle agitation or tapping of the rack before allowing to dry will decrease water droplets and their resulting spots. Air dry slides thoroughly at room temperature or at 37 °C.
VECTABOND™ Reagent solution should be prepared just prior to use. The 357 ml of solution fills a standard size horizontal staining dish and is sufficient to treat at least 500 slides. Slides can be used immediately after drying , or stored in a box at room temperature.
3. Section Adhesion:
For paraffin embedded tissue, float sections onto VECTABOND™ Reagent treated slides in a 45 °C water bath. Air dry sections thoroughly. Heat sections at 56°C for 30 minutes to affix sections well. Most tissue sections will bind well under these conditions. If there is a problem with section adhesion, heating the sections for up to 24 hours at 56 °C or for 1 hour at 75 °C will increase
adhesion. For immunohistochemical applications, temperatures above 60 °C are not recommended because loss of antigens may occur. Store slides in a box at room temperature. When needed, deparaffinize and process slides as usual. For frozen tissue, allow sections to dry on VECTABOND™ Reagent treated slides before fixation and storage. Process slides as usual.
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