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六色酪酰胺信号放大试剂盒(抗兔二抗）
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基本描述
| 英文名称 | Six color tyramide signal amplification kit (anti rabbit secondary antibody) |
|---|---|
| 储存温度 | 避光,-20°C储存 |
| 运输条件 | 超低温冰袋运输 |
| 产品介绍 |
产品介绍： 酪胺信号放大（TSA）是一种基于辣根过氧化物酶（HRP）的催化活性对靶蛋白或核酸进行高密度原位标记的酶学检测方法。原理为酪胺Tyramide的过氧化物酶反应（已标记荧光的酪胺在HRP催化H?O?下形成共价键结合位点）产生的大量酶促产物与目标蛋白的酪氨酸残基共价结合，从而使目标蛋白标记上特异的荧光。多重标记仅需在热修复法去除非共价结合的抗体后换另一种一抗、荧光素酪胺，如此往复即可。 产品参数： 荧光光谱数据：YF?488: 490/515 nm; YF?555: 555/565 nm;?YF?594: 590/617 nm; YF?640: 642/662 nm; YF?680: 681/698 nm 注意事项： 1. 使用前请将产品瞬时离心至管底，再进行后续实验。 2. 1× Tyramide Amplification Buffer首次使用后，建议小量分装，-20℃保存，避免反复冻融。 3. 为了防止出现假阴性、假阳性的结果，实验过程中需设置阴性对照和阳性对照。对于组织样品，建议对未染色的对照（不添加抗体或酪酰胺）进行成像，确定组织是否有自发荧光，排除对背景的影响。 4. 建议1:200稀释YF? Tyramide。较高的浓度可能会导致信号过强或背景高，建议从1:100到1:1000梯度稀释。 5. 可依次使用多个YF? Tyramide来标记同一样品的不同靶标，每次酪酰胺反应后需进行抗体剥离。 6. 多色标记时，依据抗原密度不同选择不同荧光素（低密度抗原选择强染料；高密度抗原选择较弱染料）。标记顺序对最终标记效果有一定影响，需自行摸索。 7. 如若做细胞样本/冰冻切片的TSA多色实验，需进行预实验判断试剂是否可用，具体步骤请参考相关文献。 应用范围： TSA、多重荧光免疫组化、肿瘤微环境分析 Product introduction： Tyramine signal amplification (TSA) is an enzymatic detection method based on the catalytic activity of horseradish peroxidase (HRP) for high-density in situ labeling of target proteins or nucleic acids. The principle is that a large number of enzymatic products produced by the peroxidase reaction of tyramine (labeled fluorescent tyramine forms a covalent bond binding site under HRP catalyzed h ? o ?) are covalently bound to the tyrosine residues of the target protein, thus making the specific fluorescence on the target protein label. The multiplex labeling only needs to change another primary antibody, fluorescein tyramine, after the non covalently bound antibody is removed by the thermal repair method, and so on. Product parameters： Fluorescence spectrum data: YF ? 488: 490 / 515 nm; YF ? 555: 555 / 565 nm; YF ? 594: 590 / 617 nm; YF ? 640: 642 / 662 nm; YF ? 680: 681 / 698 NM Matters needing attention： 1. please centrifuge the product to the bottom of the tube immediately before use, and then conduct subsequent experiments. 2.1 × After tyramide amplification buffer is used for the first time, it is recommended to pack it in small quantities and store it at -20 ℃ to avoid repeated freezing and thawing. 3. in order to prevent false negative and false positive results, negative control and positive control should be set during the experiment. For tissue samples, it is recommended to image the unstained control (without adding antibody or tyramide) to determine whether the tissue has autofluorescence and exclude the influence on the background. 4. recommend 1:200 dilution of YF ? Tyramide. Higher concentration may lead to too strong signal or high background, and gradient dilution from 1:100 to 1:1000 is recommended. 5. multiple YFS can be used in sequence ? Tyramide is used to label different targets of the same sample, and antibody stripping is required after each tyramide reaction. 6. for multicolor labeling, different fluorescens are selected according to different antigen densities (strong dyes are selected for low-density antigens; weak dyes are selected for high-density antigens). The marking sequence has a certain impact on the final marking effect, which needs to be explored by yourself. 7. if the TSA multicolor experiment of cell samples / frozen sections is done, pre experiment should be carried out to determine whether the reagent is available. Please refer to the relevant literature for the specific steps. Scope of application： TSA, multiplex fluorescence immunohistochemistry, tumor microenvironment analysis |
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