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  • Fig: Fluorescence immunohistochemical analysis of mouse-colon tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of mouse-hippocampus tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of RAT - colon tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of mouse-cortex tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of Rat-kidney tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-ATP1A1 antibody ( AWA11936 ) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA11936) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of Rat-liver tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-ATP1A1 antibody ( AWA11936 ) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA11936) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of Rat-placenta tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-ATP1A1 antibody ( AWA11936 ) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA11936) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of Rat-spleen tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-ATP1A1 antibody ( AWA11936 ) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA11936) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

荧光单标信号放大试剂(TSA-570)

货号：AWI0689

价格：￥210

规格：10T50T100T

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  • Fig: Fluorescence immunohistochemical analysis of mouse-colon tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of mouse-hippocampus tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of RAT - colon tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of mouse-cortex tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of Rat-kidney tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-ATP1A1 antibody ( AWA11936 ) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA11936) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of Rat-liver tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-ATP1A1 antibody ( AWA11936 ) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA11936) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of Rat-placenta tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-ATP1A1 antibody ( AWA11936 ) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA11936) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • Fig: Fluorescence immunohistochemical analysis of Rat-spleen tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-ATP1A1 antibody ( AWA11936 ) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA11936) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.
  • 产品概述

    • 产品介绍

    酪酰胺信号放大技术（Tyramide Signal Amplification, TSA）主要是利用酪胺的过氧化物酶反应。酪胺非活性荧光素底物在HRP和过氧化氢的作用下，会被激活产生活化荧光底物，同时形成共价键结合位点，共价结合在蛋白抗原表面或附近的酪氨酸残基上，抗原和抗体的结合部位就会有大量的酪胺荧光素沉积，使抗原位点处的荧光信号增强。

    酪胺荧光素底物-抗原酪氨酸共价稳定结合，故TSA信号不会受微波影响，可用热修复法清除第一轮与抗原非共价结合的抗体复合物，并能在抗体去除后保留与抗原相关的荧光信号。然后，再用第二种一抗进行第二轮孵育，同时更换另一种酪胺荧光素底物，多次循环反复，不同的酪胺荧光素进行标记就可实现多重免疫组化染色。

    WPS图片(1).png

    产品组成成分


    名称

    AWI0689a

    10T

    AWI0689b

    50T

    AWI0689c

    100T

    保存条件

    TSA-570 荧光染料

    0.5 ml

    2.5 ml

    5 ml

    -20℃，避光

    内源性过氧化物酶阻断剂

    0.5 ml

    2.5 ml

    5 ml

    4℃，避光

    超敏酶标山羊抗小鼠/兔 IgG 聚合物

    0.5 ml

    2.5 ml

    5 ml

    4℃，避光



    保存条件

    荧光染料 -20℃避光保存，12个月

    内源性过氧化物酶阻断剂、超敏酶标山羊抗小鼠/兔 IgG 聚合物 4℃避光保存，12个月


    实验前材料准备

    1、PBS缓冲液、5% BSA-PBS（或者其他封闭液）、柠檬酸钠抗原修复液（或其他修复液）；

    2、推荐可用于IHC-P的单克隆抗体；

    3、DAPI染色液（推荐浓度：5μg/mL 货号：AWC0298）



    操作步骤

    1、操作流程简图

    image.png

    2、详细操作步骤

    (1)石蜡切片脱蜡至水：依次将切片放入二甲苯Ⅰ 15min→二甲苯Ⅱ 15min→无水乙醇Ⅰ10min→无水乙醇Ⅱ10min→95%乙醇 5min→85%乙醇 5min→75%乙醇 5min→蒸馏水洗。

     

    (2)抗原修复：组织切片置于盛满 PH 9.0 EDTA 碱性抗原修复液或者 PH6.0 柠檬酸修复缓冲液的修复盒中于微波炉内进行抗原修复（也可以用高压 1-2min 100℃水煮 15min 95℃水浴 20min 等其他热修复方法）。中火 8min，停火 8min，转中低火 7min，此过程中应防止缓冲液过度蒸发，切勿干片。自然冷却后将玻片置于 PBS（PH7.4）中在脱色摇床上晃动洗涤 3 次，每次 5 min (修复液和修复条件根据组织类型以及抗原类型来确定）。

     

    (3)阻断内源性过氧化物：切片放入内源性过氧化物酶阻断剂，室温避光孵育 15 min，将玻片置于 PBS（PH7.4）中在脱色摇床上晃动洗涤 3 次，每次 5min。

     

    (4)BSA 封闭：切片稍甩干后用组化笔在组织周围画圈（防止抗体流走），在圈内滴加用 5% BSA-PBS（或者其他封闭液）均匀覆盖组织，(4)37℃恒温孵育 60  min（此步骤也可以使用山羊血清等封闭液进行封闭）。

     

    (5)加一抗：轻轻甩掉封闭液，在切片上滴加用抗体稀释液稀释好的的一抗 X，切片平放于避光湿盒内    4℃孵育过夜或者 37℃ 1-2h。（湿盒内加少量水防止抗体蒸发）

     

    (6)加免疫组化 HRP 二抗：玻片置于 PBS（PH7.4）中在脱色摇床上晃动洗涤 3 次，每次 5min。切片稍甩干后在圈内滴加与一抗相应种属的免疫组化 HRP 二抗覆盖组织，避光室温孵育 30-50min，PBS 洗三次。

     

    (7)荧光染料反应：TSA-570 荧光染料反应 10-15 min，PBS 洗三次。

     

    (8)DAPI 复染细胞核：玻片置于 PBS（PH7.4）中在脱色摇床上晃动洗涤 3 次，每次 5min。切片稍甩干后在圈内滴加 DAPI 染液，避光室温孵育 10min。

     

    (9)封片：玻片置于 PBS（PH7.4）中在脱色摇床上晃动洗涤 3 次，每次 5min。切片稍甩干后用抗荧光淬灭封片剂封片。

     

    (10)镜检拍照：切片于荧光显微镜/共聚焦/多通道荧光扫描仪/多光谱成像系统下观察并采集图像。


    (9)封片：玻片置于 PBS（PH7.4）中在脱色摇床上晃动洗涤 3 次，每次 5min。切片稍甩干后用抗荧光淬灭封片剂封片。


    (10)镜检拍照：切片于荧光显微镜/共聚焦/多通道荧光扫描仪/多光谱成像系统下观察并采集图像。


    染料

    激发波长

    发射波长

    DAPI

    350nm

    420nm

    TSA-480

    450nm

    480nm

    TSA-520

    490nm

    520nm

    TSA-570

    550nm

    570nm

    TSA-620

    590nm

    620nm

    TSA-690

    630nm

    690nm

    TSA-780

    750nm

    780nm


    注意事项

    1、试剂初次使用前请置于4℃解冻，解冻后于4℃短期保存，避免反复冻融，请尽快使用。

    2、为了您的安全和健康，请穿好实验服并佩戴一次性手套和口罩操作。

       3、本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品。


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