| 货号 (SKU) | 包装规格 | 是否现货 | 价格 | 数量 |
|---|---|---|---|---|
| E743397-100ml |
100ml |
期货  |
| |
| E743397-500ml |
500ml |
期货 |
|
| 产品名称 | 增强型TMB显色液(ELISA HRP显色用) |
|---|---|
| 产品介绍 |
增强型TMB显色液(ELISA HRP显色用)，英文名Enhanced TMB Chromogen Solution for ELISA或Enhanced TMB Substrate Solution for ELISA or Enhanced TMB Solution for ELISA)是一种采用了最新单一溶液TMB显色技术，通过辣根过氧化物酶(HRP)催化TMB显色，用于ELISA等的增强型显色液。本显色液也可以用于检测血液或血红蛋白等样品中的过氧化物酶含量。通常的TMB显色试剂由多个组份组成，必须在使用前进行配制，并且容易产生沉淀，使用相对不太方便，并且也容易导致检测结果不太稳定。本TMB显色液采用了最新的TMB显色技术，把所有的相关试剂全部配制在一个溶液中，仅由单一溶液组成，简化了操作步骤，并且使检测结果更加稳定可靠。TMB，即3, 3´,5 ,5´-Tetramethylbenzidine，是辣根过氧化物酶的常用底物。在辣根过氧化物酶或其他适当过氧化物酶的催化下，TMB会产生可溶性蓝色产物。蓝色产物通常可以在370nm或620-650nm测定吸光度。不同浓度辣根过氧化物酶(HRP)标记二抗使用本产品的检测效果参见图1A。图1. P0210 增强型TMB显色液(ELISA HRP显色用)的检测效果图。A. 本产品和P0209 TMB显色液(ELISA HRP显色用)的检测效果对比图。辣根过氧化物酶标记山羊抗兔IgG 用PBS稀释至图中所示浓度，各取20微升，加入200微升本产品，在5分钟时检测A370。从检测结果可知，HRP标记二抗浓度至少在0-0.02%范围内呈良好的线性关系，并且P2010的检测灵敏度比P0209约高50%。B. 本产品直接检测A370和添加终止液后检测A450的检测效果对比图。图中可见添加终止液后检测A450的检测灵敏度更高一些。实测数据会因试剂、检测仪器等的不同而存在差异，图中数据仅供参考。辣根过氧化物酶催化TMB显蓝色后可以使用阿拉丁的TMB显色终止液(450nm, 不含硫酸) 、TMB显色终止液(650nm, 无腐蚀性) 或自行配制的2M H2SO4终止反应。加入TMB显色终止液(450nm, 不含硫酸)或硫酸后，溶液呈黄色，此时可以在450nm测定吸光度，吸光度会有显著升高(图1B)；加入TMB显色终止液(650nm, 无腐蚀性)后，溶液保持蓝色不变，此时可在620-650nm测定吸光度。本试剂盒最常用于ELISA检测，也可以用于检测血液或血红蛋白等样品中的过氧化物酶含量。用于ELISA检测时，每个样品通常使用0.1毫升显色液，每100ml本产品共可以检测约1000个样品。4℃避光保存，一年有效。TMB对人体有刺激性，操作时请小心，并注意适当防护以避免直接接触人体或吸入体内。本产品为微蓝色透明溶液，如果发现TMB显色液出现混浊或颜色变成较深的蓝色，应该停止使用。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，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。 使用说明： 1.对于ELSIA检测： a.参考ELISA试剂盒的实验步骤，在与辣根过氧化物酶标记的抗体孵育后，用适当洗涤液洗涤3-5次，每次3-5分钟。 b.洗涤完毕后，去除洗涤液，加入100微升TMB显色液。 c.室温避光孵育3-30分钟或更长时间(可长达24小时)，直至显色至预期深浅。 d.直接在370nm或620-650nm测定吸光度。或加入100微升阿拉丁的TMB显色终止液(450nm, 不含硫酸) 或TMB显色终止液(650nm, 无腐蚀性) ，或自行配制2M H2SO4终止反应，随后在相应波长测定吸光度。 2.对于在96孔板内进行的其它适当检测 (例如检测组织或细胞样品内源性的过氧化物酶)： a.直接在96孔板内每孔加入10-20微升样品。 b.加入100微升TMB显色液。 c.室温避光孵育3-30分钟或更长时间(可长达24小时)，直至显色至预期深浅。 d.直接在370nm或620-650nm测定吸光度。或加入100微升阿拉丁的TMB显色终止液(450nm, 不含硫酸) 或TMB显色终止液(650nm, 无腐蚀性) ，或自行配制2M H2SO4终止反应，随后在相应波长测定吸光度。 常见问题： 1.背景显色太深。 a.如果背景(没有样品的对照)显色太深，一方面需考虑使用适当的封闭液进行封闭，例如选购适当的封闭液或使用和一抗相同来源的血清(10%)进行封闭。另一方面，请选购经过适当吸附的二抗，以减小二抗的非特异性吸附。 b.可以考虑缩短显色时间，或降低二抗浓度。另外，选择适当强度的洗涤液，或延长洗涤时间也会有所帮助。 2.没有显色或显色太弱。 a.适当提高一抗或二抗的浓度。检测二抗效果，滴一滴稀释二抗在离心管内，检测二抗是否可以被正常显色。 b.可以考虑使用更加灵敏的放大检测体系，例如使用生物素检测体系。 c.可以适当延长显色时间。 d.如果上述改进不能获得预期效果，可以考虑更换效果更好的一抗或ELISA试剂盒。The Enhanced TMB Substrate Solution for ELISA adopts the latest color development technology with a single TMB solution. The conversion of TMB into a soluble blue product is catalyzed by horseradish peroxidase (HRP). This product can also be used to assay the content of peroxidase in blood or hemoglobin samples.The traditional TMB color development reagents consists of multiple component that must be prepared freshly before use and are prone to precipitation, which is relatively inconvenient and produces inconsistent assay results easily. This product is a ready-to-use single-component solution, which greatly simplifies the procedures and makes the assay more robust and reliable.TMB, namely 3, 3',5 ,5'-Tetramethylbenzidine, is a substrate of HRP. Under the catalysis of HRP or other appropriate peroxidases, TMB generates a soluble blue product with an absorbance at 370nm or 620-650nm. Please refer to Figure 1 for the assay results of HRP-labeled secondary antibodies at different concentrations using this product.Figure 1. The assay results of HRP-labeled secondary antibodies at different concentrations using Aladdin's Enchanced TMB Substrate Solution for ELISA . A. Comparison of the assay results between the TMB Substrate Solution for ELISA and the Enhanced TMB Substrate Solution for ELISA . B. Measurement of A370 before the addition of Stop Solution and A450 after the addition of Stop Solution. As shown in the figure, the concentration of HRP-labeled secondary antibody shows good linearity in the range of 0-0.02% and the assay sensitivity of P2010 is approximately 50% higher than that of P0209. The assay sensitivity is higher after the addition of stop solution . This figure is for reference only, which may vary due to different experimental conditions.After TMB conversion into the blue product by HRP, the reaction can be stopped by the addition of Stop Solution for TMB Substrate (450nm, Sulfate Free) , Stop Solution for TMB Substrate (650nm, Non-corrosive) or home-made 2M H2SO4. After adding Stop Solution for TMB Substrate (450nm, Sulfate Free) or 2M H2SO4, the solution turns yellow and has a higher absorbance value at 450nm (Figure 1B). After adding Stop Solution for TMB Substrate (650nm, Non-corrosive), the solution remains blue and the absorbance can be measured at 620-650nm (Figure 1B).This product is most commonly used in ELISA, but can also be used to determine the content of peroxidase in blood or hemoglobin samples. When used for ELISA, each sample usually requires 0.1ml of this product. Precautions： TMB is irritating. Please take effective measures to avoid inhalation or direct contact with skin.This product is a colorless to slightly blue transparent solution. Discard it when it turns turbid or darker blue.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation. Instructions for Use： 1. For ELSIA:a. (Refer to the instructions of the ELISA kit) After incubation with HRP-labeled antibody, wash 3-5 times with an appropriate wash buffer for 3-5 minutes each.b. Remove the wash buffer and add 200μl of TMB Substrate Solution for ELISA.c. Incubate at room temperature in the dark for 3-30 minutes or longer (up to 24 hours) until the desired color intensity is achieved.d. Measure the absorbance directly at 370nm or 620-650nm. Alternatively, add 100μl of Stop Solution for TMB Substrate (450nm, Sulfate Free) (, P0215), Stop Solution for TMB Substrate (650nm, non-corrosive) (, P0217) or home-made 2M H2SO4 to terminate the reaction, then measure the absorbance.2. For other assays in 96-well plates (e.g., detection of endogenous peroxidase in cell or tissue samples):a. Add 10-20 microliters of sample directly to each well of 96-well plates.b. Add 200μl of MB Substrate Solution for ELISA.c. Incubate at room temperature in the dark for 3-30 minutes or longer (up to 24 hours) until the desired color intensity is achieved.d. Measure the absorbance directly at 370nm or 620-650nm. Alternatively, add 100μl of Stop Solution for TMB Substrate (450nm, Sulfate Free) (, P0215), Stop Solution for TMB Substrate (650nm, non-corrosive) (, P0217) or home-made 2M H2SO4 to terminate the reaction, then measure the absorbance.FAQ：1. Too dark backgrounda. If the background of negative control is too dark, attempt other appropriate blocking buffers or block with 10% serum from the same source as the primary antibody. Alternatively, use secondary antibodies that have been properly adsorbed to reduce non-specific bindings of secondary antibody.b. Reducing the color development time or the concentration of secondary antibody can be considered. Using other wash buffers with stronger strength or extending the washing time may also help.2. No color or too weak colora. Increase the concentration of the primary or secondary antibody appropriately. To test the effectiveness of the secondary antibody, add a drop of diluted secondary antibody to a centrifuge tube and check if the colour can be developed normally.b. Use a more sensitive detection system with higher amplification efficiency, such a biotin detection system.c. Extend the color development time appropriately.d. Use a primary antibody or ELISA kit with better effects. |
| 储存温度 | 2-8°C储存,避光 |
|---|---|
| 运输条件 | 冰袋运输 |
| 稳定性与储存 | 4℃避光保存，一年有效。 |
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