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Western检测

DAB辣根过氧化物酶显色试剂盒
产品编号: P0203
产品包装: 共100ml
产品价格: 219.00元
产品简介
使用说明
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产品编号 产品名称 产品包装 产品价格
P0203 DAB辣根过氧化物酶显色试剂盒 共100ml 219.00元

      DAB辣根过氧化物酶显色试剂盒(DAB Horseradish Peroxidase Color Development Kit)是一种借助辣根过氧化物酶
(HRP),用于免疫组化显色、原位杂交显色或Western、Southern、Northern、EMSA等膜显色的试剂盒。
      DAB,即3,3N-Diaminobenzidine Tertrahydrochloride,是辣根过氧化物酶的常用底物。在辣根过氧化物酶的催化下,DAB会产生棕色沉淀。该棕色沉淀不溶于水和乙醇。因此在DAB显色后,还可以使用溶于乙醇的染料进行后续染色。
      本试剂盒可以用于细胞或组织在免疫组化或原位杂交时结合的辣根过氧化物酶显色,也可以用于Western等结合有辣根过氧化物酶的膜的显色检测。同时也可以用于细胞或组织内源性的辣根过氧化物酶显色。
      用于免疫组化或原位杂交时,如果每个片子使用0.2毫升显色液,那么本试剂盒共可以检测500个片子。
包装清单:

产品编号 产品名称 包装
P0203-1 DAB显色液A 50ml
P0203-2 DAB显色液B 50ml
说明书 1份

保存条件:
      -20℃保存,一年有效。DAB显色液A和DAB显色液B均需避光保存。
注意事项:
      DAB对人体有害,操作时请小心,并注意有效防护以避免直接接触人体或吸入体内。
      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. 对于组织切片或细胞样品或膜,在与辣根过氧化物酶标记的抗体或其它形式的探针孵育后,用适当洗涤液洗涤3-5次,每次3-5分钟。对于检测内源性辣根过氧化物酶的组织或细胞样品,在适当固定后,也用适当洗涤液洗涤3-5次,每次3-5分钟。
2. 按照如下比例依次加入各溶液,混匀后即配制成DAB染色工作液:

DAB显色液A 0.5ml 5ml
DAB显色液B 0.5ml 5ml
DAB染色工作液 1ml 10ml

3. 最后一次洗涤完毕后,去除洗涤液,加入适量DAB染色工作液,确保能充分覆盖样品。
4. 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。
5. 去除DAB染色工作液,用蒸馏水洗涤1-2次即可终止显色反应。
6. 对于组织切片或细胞样品,显色反应终止后,如有必要可以用中性红染色液(neutral red staining solution)染色,以便于观察。对于膜,显色反应终止后,可以室温晾干避光保存。
常见问题:
1. 背景显色太深。
a. 在免疫组化时如果背景显色太深,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,请注意选购经过适当吸附的二抗,以减小二抗的非特异性吸附。
b. 在免疫组化时如果背景显色太深,需注意灭活内源性过氧化氢酶。可以在4体积甲醇中加入1体积3%过氧化氢,混匀后用于内源性过氧化氢酶的灭活。
c. 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. 可以考虑适当提高一抗或二抗的浓度。检测二抗效果,滴一滴稀释二抗在膜上,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,另外确定抗原修复是否对于使用的一抗是必需的。

相关产品请点击如下按钮:
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