Vega Red Chromogen Kit (Xylene Compatible)
Vega Red Chromogen Kit
For optimal performance, treat the tissue specimens with 10X Phosphatase Enhancer (Cat. No. M65) prior to introducing this chromogen mixture.
1. After incubating with the Alkaline Phosphatase detection system, rinse the slides with 1X Automation Buffer (Cat. No. M30) or washing buffer six times followed by two rinsings with distilled water.
2. Introduce chromogen mixture and incubate for 10 to 20 minutes or until the desired level of staining has been achieved.
3. Wash the tissue slides twice with distilled water.
4. Apply the aqueous hematoxylin counterstain (Cat. No. M10) to slides.
5. Incubate the slide for 20 to 30 seconds at room temperature.
6. Remove the aqueous hematoxylin and wash the tissue slides 2-3 times with distilled water.
7. Rinse the slides once with 1X Automation Buffer or PBS (pH 7.4) and once with distilled water.
8. Dehydrate the slide by passing them through graded alcohols (70%, 95%, 100%) and xylene or an equivalent organic solvent.
Vega Red Chromogen is a new alkaline phosphatase chomogen designed to be used when dehydration of the tissue section specimens is desirable. This chromogen will not dissolve in alcohol nor xylene. The final appearance characteristics of this chromogen are similar to those produced by Fast Red; however, Vega Red is highly stable to organic solvents. When used with hematoxylin counterstain, it produces the high contrast that blue and red colors hold. Although designed to be used in cases of dehydration of specimens, it performs equally well with aqueous mounting media such as Crystal/Mount and Gel/Mount.
Refrigerate at 4°C. Do not freeze.
REAGENTS SUPPLIED: Reagent 3A, Reagent B (Vega Red), Reagent C
Amount supplied: 10 mL Amount supplied: 20 Tablets Amount suppled: 5.0 mLPREPARATION OF WORKING SOLUTION:1. To 4 mL of deionized or distilled water, add 10 drops of reagent 3A, and mix.
2. Add 1 tablet of Vega Red Chromogen (Reagent 3B). Vortex vigorously or mix occasionally for 5 minutes or until the tablet is completely dissolved.
3. Add 5 drops of Reagent 3C. Mix well.Working solutions should be used within two hours of preparation. Discard unused portions according to local regulations.MOUNTING OF SPECIMENS:Vega Red Chromogen can be dehydrated and mounted in organic solvent-based mounting media such as Clarion (Cat. No. M05) or Permount.