peroxidase staining assay
peroxidase staining assay
The peroxide E in the cell decomposes hydrogen peroxide to release O atoms, which oxidize benzidine and change it to benzidine orchid, causing the cell plasma to show orchid-black particles, This experiment is from the Mudanjiang Medical College undergraduate 5-year lab guide for testing majors
Operation method
peroxidase staining assay
Principle
Peroxide E in the cell breaks down hydrogen peroxide to release O atoms that oxidize benzidine to benzidine blue, giving an orchid-black granule in the cytoplasm
Materials and Instruments
Benzidine Basic magenta Hydrogen peroxide solution Distilled water Move I. Experimental reagents: Caveat The smeared blood and bone marrow slices should be freshly stained and kept for a few days at most; slices that have been fixed should not be re-stained because the activity of the alcohol has been destroyed, otherwise false positives occur. For more product details, please visit Aladdin Scientific website.
1. benzidine 0.3g basic magenta 0.3g
Sodium nitroso ferrocyanide saturated aqueous solution lml (360 mg / ml) 95% diol, 100 ml of this solution is stored in a brown glass bottle, available for 8-10 months 2, 3% hydrogen peroxide solution 0.3 ml before use to re-formulate
Distilled water 25ml
Second, the experimental method:
1. Prepare blood tablets for drying as usual
2. Add 10 drops of ① solution and dye for 1-1.5 minutes.
3. Do not pour away the ① solution, then add 0.5ml of ② solution and dye for 4-5 minutes.
4. Rinse with distilled water for 0.5 to 1 minute.
5. when the glass slice is still wet, 95% ethanol decolorization until the red color fades. 6. rinse with water, leave to dry, Ricci dye re-dyeing for 20 min.
6. Judgement of the results: when positive, there are blue-black particles in the cytoplasm, and the degree of POX positivity is generally divided into weak positivity (the positive particles are small and scattered), positivity (the particles are medium-sized and a small number of them are distributed in clusters), and strong positivity (the particles are large and distributed in clusters).