计算溶液所需的质量、体积或浓度。
普鲁士蓝染色液 (DAB增强法)
有货
为什么选择此级别
BioReagent,生物染色剂,用于显微镜 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存,避光,室温。低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
含铁血黄素 (Hemosiderin)是一种血红蛋白源性色素，为金黄色或棕黄色颗粒，因其含铁，且为金黄色，故称为含铁血黄素。当红细胞被巨噬细胞吞噬后，在溶酶体酶的作用下，血红蛋白被分解为不含铁的橙色血质和含铁的含铁血黄素。含铁血黄素染色又称为Perls普鲁士蓝反应 (Prussian blue reaction) ，用于显示局部组织内的各种出血性病变，常见于吞噬细胞内，可以很好地区分含铁血黄素与其他色素。
普鲁士蓝染色液 (DAB增强法)其染色原理是组织中的三价铁离子从蛋白质中被稀盐酸分离出来，与亚铁氰化钾形成一种不溶解的蓝色的亚铁氰化铁沉淀而显示出组织中的高含量铁元素，当组织中的铁元素含量极少时，该染色不能显示出蓝色定位铁元素，加入DAB后，与亚铁氰化铁沉淀发生氧化还原反应形成棕色化合物，增强了检测的特异性，特别适用于组织铁含量较少的组织。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
| P1508698 | Component | 5×50 mL | Storage |
| P1508698A | Perls Stain A | 25 mL | RT. Store in the dark. |
| P1508698B | Perls Stain B | 25 mL | RT. Store in the dark. |
| P1508698C | DAB 试剂 | 1 支 | 2-8℃. Store in the dark. |
| P1508698D | DAB 氧化剂 | 1 支 | 2-8℃. Store in the dark. |
| P1508698E | DAB buffer | 2×50 mL | RT. |
| P1508698F | 苏木素染色液 | 50 mL | RT. Store in the dark. |
| P1508698G | 酸性乙醇分化液 | 50 mL | RT. |
1. 将A: Perls Stain A和B: Perls Stain B等量混合即为Perls Stain，不宜提前配制。
2. 取1支C: DAB试剂加入到30ml E: DAB buffer中，充分溶解待用，为DAB溶液；取1μl D: DAB氧化剂加入到10ml E: DAB buffer中，充分混合，为DAB氧化工作液；临用前将DAB溶液与DAB氧化工作液等比例混合，即为DAB Stain。
自备材料：
固定液：10%中性福尔马林、4%多聚甲醛等、系列乙醇、蒸馏水、二甲苯、5%草酸
操作步骤(仅供参考)：
临用前配制Perls Stain和DAB Stain：
1. 配制Perls Stain: 将Perls Stain A和Perls Stain B等量混合即为Perls Stain，不宜提前配制。
2. 配制DAB Stain: 取1支DAB试剂加入到30ml DAB buffer中，充分溶解待用，为DAB溶液；取1μl DAB氧化剂加入到10ml DAB buffer中，充分混合，为DAB氧化工作液；临用前将DAB溶液与DAB氧化工作液等比例混合，即为DAB Stain。
(一)石蜡切片染色
1、组织固定于10%中性福尔马林固定液，常规脱水包埋。
2、切片厚度4μm，常规二甲苯或浸蜡脱蜡透明液脱蜡至水，蒸馏水水洗1min。
3、切片入Perls Stain (见注意事项2)浸染15-30min。
4、蒸馏水充分冲洗2-5min。
5、滴加DAB Stain染色5-10min，显微镜下控制显色程度，倾去染液用DAB buffer浸洗 1 次，蒸馏水洗 3 次。
6、苏木素染色1-2min，自来水洗，酸性乙醇分化液分化2-5s，自来水洗5-10min。
7、常规脱水，二甲苯或浸蜡脱蜡透明液透明，中性树胶封固。
8、显微镜镜检，图像采集分析。
(二)冰冻切片染色
1、无需脱蜡，直接迅速用蒸馏水冲洗2-3min。
2、染色、封固步骤同石蜡切片的染色步骤，时间可以相应缩短。
染色结果：
含铁血黄素或三价铁: 棕褐色
细胞核: 浅蓝色
背景或其他组织: 浅棕色或无色
阴性对照(可选)：
取连续切片脱蜡至水；入5%草酸孵育2-6h，经Perls Stain，其余步骤同上；结果为阴性。
注意事项：
1、切片脱蜡应尽量干净，组织固定常采用10%中性福尔马林，经普通福尔马林长期固定后，组织会有损伤；避免使用酸性固定剂，铬酸盐处理也会妨碍铁的保存。
2、整个操作过程中容器要干净，避免用金属铁制品，洗切片和容器时以蒸馏水为宜，因普通水内含铁质；Perls Stain染色时应根据样本情况调整着色时间。
3、所有切片都应使用同一个阳性对照切片，选择适合的对照非常重要，尸检肺组织是一个很好的对照，包含相当数量的铁阳性巨噬细胞 (心衰细胞)。
4、DAB溶液配制后应-20℃保存，可短期4℃保存；DAB氧化工作液可短期4℃保存，不宜久置，应经常更换新液。如染色效果不好，可增加DAB氧化工作液中氧化剂的加入量。
5、在DAB的显色过程中，每隔约 5min 需要在显微镜下观察显色效果，由普鲁士蓝染出的弱阳性 (即浅蓝色)被DAB替代形成棕色或者无蓝色的部位变成棕色即可。
6、为了您的安全和健康，请穿实验服并戴一次性手套操作。
7、试剂开封后请尽快使用，以防影响后续实验效果。
Hemosiderin is a hemoglobin-derived pigment that exists as golden yellow or brownish yellow granules. It is named hemosiderin due to its iron content and golden color. When red blood cells are phagocytosed by macrophages, hemoglobin is decomposed into iron-free hemofuscin and iron-containing hemosiderin under the action of lysosomal enzymes. Hemosiderin staining, also known as the Perls Prussian blue reaction, is used to visualize various hemorrhagic lesions in local tissues. It is commonly found in phagocytes and can effectively distinguish hemosiderin from other pigments.
The Prussian Blue Staining Kit (DAB-Enhanced Method) works on the following principle: trivalent iron ions in tissues are dissociated from proteins by dilute hydrochloric acid, and then react with potassium ferrocyanide to form an insoluble blue iron ferrocyanide precipitate, thereby indicating high iron content in tissues. When the iron content in tissues is extremely low, this staining method cannot produce blue precipitate for iron localization. The addition of DAB triggers a redox reaction with the iron ferrocyanide precipitate, forming a brown compound, which enhances the specificity of detection. This method is particularly suitable for tissues with low iron content. This reagent is for research use only and not intended for clinical diagnosis or other purposes.
| P1508698 | Component | 5×50 mL | Storage |
| P1508698A | Perls Stain A | 25 mL | RT. Store in the dark. |
| P1508698B | Perls Stain B | 25 mL | RT. Store in the dark. |
| P1508698C | DAB Reagent | 1 EA | 2-8℃. Store in the dark. |
| P1508698D | DAB Oxidant | 1 EA | 2-8℃. Store in the dark. |
| P1508698E | DAB Buffer | 2×50 mL | RT. |
| P1508698F | Hematoxylin Staining Solution | 50 mL | RT. Store in the dark. |
| P1508698G | Acid Alcohol Differentiating Solution | 50 mL | RT. |
Perls Stain Preparation: Mix equal volumes of Perls Stain A (P1508698A) and Perls Stain B (P1508698B). Do not prepare in advance.
DAB Stain Preparation: Add 1 vial of DAB Reagent (P1508698C) into 30 mL of DAB Buffer (P1508698E) and dissolve completely to obtain the DAB solution. Add 1 μL of DAB Oxidant (P1508698D) into 10 mL of DAB Buffer (P1508698E) and mix thoroughly to obtain the DAB oxidation working solution. Immediately before use, mix the DAB solution and DAB oxidation working solution in equal proportions to prepare the DAB Stain.
Self-Prepared Materials
Fixatives: 10% neutral formalin, 4% paraformaldehyde, etc. Graded ethanol, distilled water, xylene, 5% oxalic acid
Operating Procedures (For Reference Only)
Prepare Perls Stain and DAB Stain Before Use:
1. Perls Stain Preparation: Mix equal volumes of Perls Stain A and Perls Stain B. Do not prepare in advance.
2. DAB Stain Preparation: Add 1 vial of DAB reagent into 30 mL of DAB buffer and dissolve completely for later use as the DAB solution; add 1 μL of DAB oxidant into 10 mL of DAB buffer and mix thoroughly as the DAB oxidation working solution; mix the DAB solution and DAB oxidation working solution in equal proportions immediately before use to obtain the DAB Stain.
(1) Staining of Paraffin Sections
1. Fix tissues in 10% neutral formalin fixative, followed by routine dehydration and embedding.
2. Cut sections at a thickness of 4 μm. Deparaffinize to water using routine xylene or paraffin-immersed clearing solution, then rinse with distilled water for 1 min.
3. Immerse sections in Perls Stain (see Note 2) for 15-30 min.
4. Rinse thoroughly with distilled water for 2-5 min.
5. Add DAB Stain dropwise and stain for 5-10 min. Monitor color development under a microscope. Pour off the staining solution, rinse once with DAB buffer, and wash 3 times with distilled water.
6. Stain with hematoxylin for 1-2 min, rinse with tap water, differentiate with acid alcohol differentiating solution for 2-5 s, then rinse with tap water for 5-10 min.
7. Perform routine dehydration, clear with xylene or paraffin-immersed clearing solution, and mount with neutral balsam.
8. Examine under a microscope, then collect and analyze images.
(2) Staining of Frozen Sections
1. No deparaffinization required; directly rinse rapidly with distilled water for 2-3 min.
2. Follow the same staining and mounting steps as for paraffin sections, with appropriately shortened incubation times.
Staining Results
Hemosiderin or trivalent iron: Brownish
Cell nucleus: Light blue
Background or other tissues: Light brown or colorless
Negative Control (Optional)
Take serial sections, deparaffinize to water; incubate in 5% oxalic acid for 2-6 h, stain with Perls Stain, and proceed with the remaining steps as described above; the expected result is negative.
Precautions
1. Ensure thorough deparaffinization of sections. Tissues are usually fixed in 10% neutral formalin; prolonged fixation in ordinary formalin may cause tissue damage. Avoid using acidic fixatives; chromate treatment can also interfere with iron preservation.
2. Keep all containers clean throughout the operation; avoid using iron metal utensils. Use distilled water for rinsing sections and washing containers, as tap water contains iron ions. Adjust the staining time of Perls Stain according to sample conditions.
3. Use the same positive control section for all test sections; selecting an appropriate control is critical. Autopsied lung tissue is an excellent control, as it contains a considerable number of iron-positive macrophages (heart failure cells).
4. Store the prepared DAB solution at -20℃, or at 4℃ for short-term use; the DAB oxidation working solution can be stored at 4℃ for short periods, but should not be left standing for extended times and must be replaced regularly. If staining results are unsatisfactory, increase the amount of oxidant added to the DAB oxidation working solution.
5. During DAB color development, observe the staining under a microscope every 5 min. Stop the reaction when the weak positive signal (light blue) from Prussian blue staining is replaced by DAB to form a brown color, or when the non-blue areas turn brown.
6. For your safety and health, wear a lab coat and disposable gloves during operation.
7. Use the reagents as soon as possible after opening to avoid affecting subsequent experimental results.
规格
别名
普鲁士蓝染色液
英文别名
Prussian Blue Staining Solution
规格或纯度
BioReagent,用于显微镜,生物染色剂
稳定性与储存
各组分在相应的储存条件下保质期1年。
英文名称
Prussian Blue Staining Solution (DAB Enhancement Method)
储存条件
2-8°C储存,避光,室温
运输条件
低温运输
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
化学和物理性质
敏感性
Light-sensitive
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 E2612522 |
分析证书 | P1508698 |
| ZJ26F0131584 |
分析证书 | P1508698 |
| ZJ26F0131583 |
分析证书 | P1508698 |
| ZJ26F0131582 |
分析证书 | P1508698 |
| ZJ26F0131581 |
分析证书 | P1508698 |
| ZJ26F0131580 |
分析证书 | P1508698 |
| ZJ26F0131579 |
分析证书 | P1508698 |
| ZJ26F0131578 |
分析证书 | P1508698 |
技术文档和文章
Principles, Classification, and Clinical Laboratory Applications of Hematologic Cytochemical Staining
目录： 技术文章
Application Differences Among Highman, Bennhold, Puchtler, and Modified Methods in Congo Red Staining for Amyloid
目录： 技术文章
溶液计算器
常见问题
什么是「生物染色剂」？
「生物染色剂」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
什么是「BioReagent」？
「BioReagent」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
本产品应如何储存？
请于2–8 °C、避光条件下保存。本品对光敏感，请保存在原始的避光或棕色容器中。
本产品如何运输？
本产品采用湿冰冷藏运输。收货后请立即拆包，并按上述储存条件保存。
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