总一氧 化氮检测试剂盒
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总一氧 化氮检测试剂盒

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  • 碧云天(beyotime)已认证
  • S0024
  • 2025年11月10日
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    总一氧 化氮检测试剂盒(Total Nitric Oxide Assay Kit, 即Nitrate/Nitrite Assay Kit)采用了硝酸盐还原酶(Nitrate reductase)还原硝酸盐(nitrate)为亚硝酸盐(nitrite),然后通过经典的Griess reagent检测亚硝酸盐, 从而测定出总一氧 化氮。一氧 化氮本身极不稳定,在细胞内很快代谢为硝酸盐(Nitrate)和亚硝酸盐(Nitrite),通过上述方法检测出硝酸盐和亚硝酸盐的总量,就可以推算出总的一氧 化氮的量。
    本试剂盒采用了NADPH依赖性硝酸盐还原酶(NADPH dependent nitrate reductase)。高浓度的NADPH会干扰后续的检测,消除NADPH的一种常用方法就是使用Lactate dehydrogenase(LDH)清除NADPH。本试剂盒采用了LDH清除NADPH的方法,使检测结果更加准确。
    对亚硝酸盐的检测下限达到2微摩尔/升,在2-80微摩尔/升的范围内有很好的线性关系。浓度过高的样品可以适当稀释后再进行检测。
    样品范围广,可以检测细胞裂解液、组织裂解液、细胞或组织的培养液、血清、血浆或尿液等中一氧 化氮的含量。酚红和10%血清对测定无 明显干扰。
    样品需要量少。根据样品中一氧 化氮的浓度不同,仅需0-60微升样品。
    检测速度快,仅需约80分钟即可完成检测。
    本试剂盒采用了间接的一氧 化氮检测方法,如需检测细胞内实际的一氧 化氮水平,可以采用碧云天生产的DAF-FM DA(NO荧光探针)(S0019)。
    包装清单:

    产品编号产品名称包装
    S0024-1样品稀释液15ml
    S0024-2NADPH5mg
    S0024-3FAD1.1ml X2
    S0024-4Nitrate Reductase1ml
    S0024-5LDH Buffer1.1ml X2
    S0024-6LDH1ml X2
    S0024-7NaNO2(1M)1ml
    S0024-8Griess Reagent I12ml
    S0024-9Griess Reagent II12ml
    说明书1份

    保存条件:
    -20℃保存,一年有效。NADPH,Nitrate Reductase,NaNO2 (1M),Griess Reagent I和Griess Reagent II需避光保存。NADPH配制成溶液后必须分装并-70℃保存。
    注意事项:
    RPMI 1640等含有较高浓度硝酸盐的培养液容易对本试剂盒的检测产生干扰,请尽量避免。必需使用RPMI 1640培养液时,在检测一氧 化氮前必需先换成其他适当培养液例如DMEM、MEM、F12等,或换成HBSS或PBS等。
    推荐选购碧云天生产的细胞与组织裂解液(一氧 化氮检测用)(S3090)用于细胞或组织样品的裂解。也可以自行选择适当方式制备细胞或组织样品的裂解液用于总一氧 化氮的检测,但需注意自行选择的方式可能会干扰试剂盒检测时的酶反应而影响检测效果。检测细胞或组织的培养液、血清、血浆或尿液中的总一氧 化氮含量时,不需要选购S3090。S3090裂解获得的样品还可以用于蛋白浓度的检测以及Western印迹检测,但需注意S3090中不含蛋白酶和磷酸酯酶抑制剂。
    检测反应必须避光进行。
    由于检测过程中有还原反应,凡是影响还原反应的氧化或还原试剂要注意避免,例如常用的还原剂DTT和巯基乙醇。
          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。
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    总一氧 化氮检测试剂盒
    ¥1850