碧云天生物技术-TMB显色液(ELISA HRP显色用)(P0209-500ml)

试剂> 多肽与蛋白> 蛋白检测> ELISA

TMB显色液(ELISA HRP显色用)

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产品编号: P0209-500ml

产品包装:500ml

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产品编号	产品名称	产品包装	产品价格
P0209-100ml	TMB显色液(ELISA HRP显色用)	100ml	243.00元
P0209-500ml	TMB显色液(ELISA HRP显色用)	500ml	898.00元

      TMB显色液(ELISA HRP显色用)(TMB Chromogen Solution for ELISA, TMB Substrate Solution for ELISA or 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1。

图1. 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图。辣根过氧化物酶标记山羊抗兔IgG (A0208)用PBS稀释至图中所示浓度，各取20微升，加入200微升本产品，分别在5分钟和30分钟时检测A370。从结果可知，5分钟时，HRP浓度在0-0.1%范围内呈较好的线性关系；30分钟时，HRP浓度在0-0.06%范围内呈较好的线性关系。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显蓝色后可以使用碧云天的TMB显色终止液(450nm, 不含硫酸) (P0215)、TMB显色终止液(650nm, 无腐蚀性) (P0217)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；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。两种TMB显色终止液加入后的吸光度检测和实拍显色效果图参考图2和图3。

图2. 不同浓度辣根过氧化物酶加入本产品后，再使用TMB显色终止液(450nm, 不含硫酸) (P0215)终止反应后的检测效果图。左图为不同浓度的HRP标记的二抗(A0208 辣根过氧化物酶标记山羊抗兔IgG (H+L))，加入本产品室温反应5分钟后A370的吸光值，及加入TMB显色终止液(450nm, 不含硫酸) A450的吸光值的线性图。右图为两者的实拍显色效果图。图中数据仅供参考，实际的检测效果可能会略有不同。

图3. 不同浓度辣根过氧化物酶加入本产品后，再使用TMB显色终止液(650nm, 无腐蚀性) (P0217)终止反应后的检测效果图。左图为不同浓度的HRP标记的二抗(A0208 辣根过氧化物酶标记山羊抗兔IgG (H+L))，加入本产品室温反应5分钟后A370的吸光值，及加入TMB显色终止液(650nm, 无腐蚀性) A650的吸光值的线性图。右图为两者的实拍显色效果图。图中数据仅供参考，实际的检测效果可能会略有不同。

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。
包装清单：

产品编号	产品名称	包装
P0209-100ml	TMB显色液(ELISA HRP显色用)	100ml
P0209-500ml	TMB显色液(ELISA HRP显色用)	500ml
—	说明书	1份

保存条件：
      4℃避光保存，一年有效。
注意事项：
      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。
      本产品为无色至微蓝色透明溶液，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微升碧云天的TMB显色终止液(450nm, 不含硫酸) (P0215)或TMB显色终止液(650nm, 无腐蚀性) (P0217)，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孔板内进行的其他适当检测 (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微升碧云天的TMB显色终止液(450nm, 不含硫酸) (P0215)或TMB显色终止液(650nm, 无腐蚀性) (P0217)，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，或使用P0210增强型TMB显色液(ELISA HRP显色用)。
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。

相关产品：

产品编号	产品名称	包装
P0209-100ml	TMB显色液(ELISA HRP显色用)	100ml
P0209-500ml	TMB显色液(ELISA HRP显色用)	500ml
P0210-100ml	增强型TMB显色液(ELISA HRP显色用)	100ml
P0210-500ml	增强型TMB显色液(ELISA HRP显色用)	500ml
P0215-100ml	TMB显色终止液(450nm, 不含硫酸)	100ml
P0215-500ml	TMB显色终止液(450nm, 不含硫酸)	500ml
P0217-100ml	TMB显色终止液(650nm, 无腐蚀性)	100ml

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