国内的胰岛素Elisa试剂盒，真的不好吗？

千姿百态

国内的胰岛素Elisa试剂盒，真的不好吗？
原文地址：https://www.zhihu.com/question/68221201

大力水手

Q1：选择哪个货号？超敏，高敏和广域胰岛素试剂盒？
答：根据检测胰岛素浓度范围，分为超敏，高敏和广域系列。它们的质量一样优秀，只是检测范围不同。在不同的实验条件下, 各种样品间的胰岛素含量可以有上千倍的差别, 目前市面上还没有一款试剂盒可以满足如此大的检测范围要求, 因此根据具体的实验情况来选择合适的试剂盒显得尤为重要, 以下建议可供参考:
① 正常小鼠或大鼠(wild type) 禁食 (fasting) 超过8小时后其胰岛素含量会较低，建议使用超敏试剂盒 (货号：MS100/RT100.检测范围: 0.05 to 2 ng/ml)。
② 小鼠或大鼠在饱食状态或禁食不超过6小时，建议使用高敏试剂盒 (货号：MS200/RT200.检测范围: 0.2 to 6.9 ng/ml)。
③ 在胰岛(Islet)或β细胞分泌实验中，样品的胰岛素含量较高，建议使用广域试剂盒 (货号：MS300/RT300.检测范围: 1 to 40 ng/ml)。另外，此类实验通常会出现胰岛素含量很高的样品，使用者需要先通过预实验来确定样品的稀释倍数以取得较好的测定结果。


Q2：实验结果必须用log-log 分析计算吗？
答：不是。线性回归分析，取对数后，线性回归分析，或者多项式或者4参数法分析都是可以的。ELISA结果计算，常用4参数方法。在线软件：https://www.mycurvefit.com/或本地软件：origin9.0
Q3：关于葡萄糖刺激胰岛素分泌glucose-stimulated insulin secretion (GSIS) 实验有哪些注意事项？
答：根据经验，有以下建议：
① 建议小鼠年龄12-20周。禁食时间不宜过长。建议禁食时间8 小时。
② 腹腔注射葡萄糖后，10分钟可以看到胰岛素明显上升。口服葡萄糖，时间会相应延长。
③ 尾部取血，避免溶血样本，不取第一滴血。尽量取顺流出的血样。取血动作一般在30秒内完成。避免溶血标本。分离血清后，为保证样品干净，10,000rpm再次离心2-3分钟。
④ Insulin & glucose 先升高，再降低。但是Insulin 浓度维持的短，Glucose 维持时间长。
⑤ High fat diet mice insulin 升高明显，有时成倍增加。但是Lean mice insulin 一般只增加20-30%。
⑥ 标准操作是在有水平震荡仪（600rpm）中孵育。在没有水平震荡仪的情况，请务必确保混匀sample。同时建议增加50%的孵育时间(in the first step)来增强反应信号。具体时间请参考对应货号的说明书。
⑦ 该实验操作较繁琐，一般需要2~3个人配合完成。
Q4：用高脂饮食诱导C57小鼠，造成胰岛素抵抗模型，大概需要多少时间？胰岛素抵抗模型建成的指标是哪些？
答：小鼠6或8 周大起诱导8 至12 周，注射胰岛素看血糖的降幅。在打完胰导素后二十分钟取第一个血样测血糖与起点比，一般對照組血糖可降50％左右，抵抗組如果小于20％可认为成功建立。
Q5：胰岛素单位如何换算？
答：1ng/ml *172.18=1 pmol/L；1 µIU/ml=0.0348 ng/ml
Q6：胰岛素(insulin)试剂盒检测结果是否包括胰岛素原（proinsulin）？
答：检测结果包括 proinsulin 和insulin。抗体对识别是两者的共同区域。
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检验医师

你听谁说的，国内的牌子咋就不好了，你买进口的说不定也是国内贴标邮过来的