过氧化物酶 (Peroxidase，简称POX或MPO) 是由微生物或植物所产生的一类能催化很多反应的以过氧化氢为电子受体催化底物氧化的酶氧化还原酶，主要存在于细胞的过氧化物酶体中，以铁卟啉为辅基，可催化过氧化氢氧化酚类和胺类化合物，具有消除过氧化氢和酚类、胺类毒性的双重作用。过氧化物酶还是细胞内重要的内源性活性氧清除剂，从而与植物抗病性有密切关系。
过氧化物酶染色液 (联苯胺法) 是ICSH推荐采用的POX染色液，其原理是细胞内的过氧化物酶能将无色的3,3-二氨基联苯胺(DAB)的氢原子传递给过氧化氢，使前者催化成有色染料沉积在细胞质中的POX所在部位。该染液主要由乙酸萘酚溶液、坚牢蓝RR染色液组成，混合后使用，可用于血液、骨髓或细胞涂片过氧化物酶染色，POX活性部位呈棕黄色。
P1518605
| Component
| 4×10mL
| 4×20mL
| Storage
|
P1518605A
| BFA 固定液
| 10mL
| 20mL
| 2-8℃. Store in the dark.
|
P1518605B
| DAB 染色液
| 10mL
| 20mL
| 2-8℃. Store in the dark.
|
P1518605C
| DAB 氧化剂
| 100μL | 200μL
| 2-8℃. Store in the dark.
|
P1518605D
| WG 染色液
| 10mL
| 20mL
| RT. Store in the dark.
|
P1518605E
| WG 缓冲液
| 10mL
| 20mL
| RT
|
临用前，按B:C=1000:1比例混合，即为 POX 孵育液，即配即用。
自备材料：
1、载玻片
2、显微镜
操作步骤（仅供参考）：
1、血液、骨髓或细胞涂片滴加预冷的BFA固定液，固定，稍水洗。
2、滴加配制好的POX孵育液，室温(20～25°C)避光孵育，水洗。
3、滴加WG染色液，孵育。
4、直接滴加等量WGbuffer，染色。
5、水洗、晾干、镜检。
染色结果：
POX活性部位：棕黄色
细胞核：蓝色
粒细胞系除早期原粒细胞阴性外，分化好的原粒细胞以下阶段细胞随细胞成熟而阳性反应增强，衰老中性粒细胞反应程度减弱单核细胞系弱阳性，淋巴细胞系为阴性。浆细胞及巨核细胞均为阴性。嗜酸性粒细胞和Auer小体呈强阳性反应。
阳性反应强度的判断：
阴性：无颗粒
弱阳性：颗粒小，分布稀疏
阳性：颗粒略粗，分布密集
强阳性：颗粒粗大，呈蓝黑色，充满胞浆
临床意义：
1、急性粒细胞白血病晚期的原粒细胞呈阳性，颗粒较少且大。
2、急性单核白血病细胞呈阴性或弱阳性，颗粒小且稀疏。
3、单核急性白血病呈阴性或弱阳性。
4、急性早幼粒白血病呈强阳性，某些早幼粒细胞呈阳性，恶性组织细胞呈阴性。
5、急性淋巴细胞白血病呈阴性。
注意事项：
1、血液或骨髓涂片应新鲜，薄厚适宜，及时固定，否则会影响酶的活性。
2、POX孵育液易失效或降低阳性强度，即配即用，不宜久置。
3、样本在未染色前切勿接触氧化剂类物质，以免细胞内的过氧化物酶被抑制。
4、每次染色时，应采取健康人末梢血或骨髓涂片作为阴性对照。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
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.
J2613214
危险品化学品经营许可证（带存储）