Peroxidase (POX or MPO for short) is a class of oxidoreductases produced by microorganisms or plants, which catalyze various reactions and use hydrogen peroxide as an electron acceptor to oxidize substrates. It is mainly located in the peroxisomes of cells, with iron porphyrin as the prosthetic group. It can catalyze the oxidation of phenols and amines by hydrogen peroxide, exerting a dual function of eliminating hydrogen peroxide and detoxifying phenols and amines. Peroxidase is also an important endogenous reactive oxygen species scavenger in cells, and is therefore closely related to plant disease resistance.
Peroxidase Staining Solution (Benzidine Method) is the POX staining solution recommended by ICSH. Its principle is that peroxidase in cells can transfer hydrogen atoms from colorless 3,3'-diaminobenzidine (DAB) to hydrogen peroxide, catalyzing the formation of a colored dye that deposits at the sites of POX in the cytoplasm. This staining solution mainly consists of naphthol acetate solution and Fast Blue RR staining solution, which are mixed before use. It can be used for peroxidase staining of blood, bone marrow or cell smears, and the POX-active sites appear yellowish-brown.
Product Components and Storage Conditions:
P1518605
| Component
| 4×10mL
| 4×20mL
| Storage
|
P1518605A
| BFA Fixative Solution
| 10mL
| 20mL
| 2-8℃. Store in the dark.
|
P1518605B
| DAB Staining Solution
| 10mL
| 20mL
| 2-8℃. Store in the dark.
|
P1518605C
| DAB Oxidizing Solution
| 100μL | 200μL
| 2-8℃. Store in the dark. |
P1518605D
| Wright‑Giemsa Staining Solution
| 10mL
| 20mL
| RT. Store in the dark.
|
P1518605E
| Wright‑Giemsa Buffer
| 10mL
| 20mL
| RT |
Immediately before use, mix at a ratio of B:C = 1000:1 to prepare the POX incubation solution. Use freshly prepared.
Materials Required (User-supplied):
1. Glass slides
2. Microscope
Procedure (For reference only):
1. Apply pre-chilled BFA Fixative Solution to blood, bone marrow or cell smears for fixation, then rinse briefly with water.
2. Add freshly prepared POX Incubation Solution, incubate at room temperature (20–25 °C) in the dark, then rinse with water.
3. Apply WG Staining Solution and incubate.
4. Add an equal volume of WG Buffer directly and stain.
5. Rinse with water, air dry, and examine microscopically.
Staining Results:
POX-active sites: Yellowish-brown
Cell nuclei: Blue
In the granulocytic series, early myeloblasts are negative, while cells at stages beyond differentiated myeloblasts show an increasingly positive reaction with maturation. The reaction intensity decreases in senescent neutrophils. The monocytic series is weakly positive. The lymphocytic series is negative. Plasma cells and megakaryocytes are both negative.Eosinophils and Auer rods show a strongly positive reaction.
Interpretation of Positive Reaction Intensity:
Negative: No granules
Weakly positive: Small granules, sparsely distributed
Positive: Moderately coarse granules, densely distributed
Strongly positive: Coarse granules, blue‑black, filling the cytoplasm
Clinical Significance:
1. Myeloblasts in advanced acute myeloid leukemia are positive, with few and large granules.
2. Cells in acute monocytic leukemia are negative or weakly positive, with small and sparse granules.
3. Monocytic acute leukemia is negative or weakly positive.
4. Acute promyelocytic leukemia is strongly positive; some promyelocytes are positive, while malignant histiocytes are negative.
5. Acute lymphoblastic leukemia is negative.
Precautions:
1. Blood or bone marrow smears should be fresh, properly thin, and fixed promptly; otherwise, enzyme activity will be affected.
2. POX incubation solution is prone to inactivation or reduced staining intensity. Prepare fresh immediately before use and do not store.
3. Unstained samples must not contact oxidizing agents, so as to avoid inhibition of intracellular peroxidase.
4. A peripheral blood or bone marrow smear from a healthy individual should be used as a negative control in each test.
5. For your safety and health, please wear a lab coat and disposable gloves during operation.
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