英文名称 :Total Protein Thiol Content Assay Kit
产品包装 :盒装
产品规格 :50T/48S
储存条件 :2-8℃
检测方法 :可见分光光度法
有 效 期 : 6个月
产品简介 :巯基的存在使得蛋白质能够进行二硫键的形成,从而维持分子的稳定性和功能性。此外,巯基还参与到氧化还原反应中,具有重要的生物学作用。在细胞内,巯基含量的变化与多种疾病的发生和进展密切相关,因此巯基也成为了生物医学领域中的重要研究对象。本试剂盒测定的是蛋白质二硫键断裂产生的巯基和本身游离巯基的总和。
还原剂会使二硫键裂解生成巯基,巯基会发生亲核反应,即巯基与5,5’-二硫代-双-硝基苯甲酸(DTNB)反应,生成黄色化合物,在412nm处有吸收峰,据此可以计算蛋白质总巯基含量。
产品组成 :
试剂名称
|
规格
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保存条件
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提取液一
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液体 40 mL×1 瓶
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2-8℃保存
|
提取液二
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液体 40 mL×1 瓶
|
2-8℃保存
|
试剂一
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液体 125 mL×1 瓶
|
2-8℃保存
|
试剂二
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液体 20 mL×1 瓶
|
2-8℃保存
|
试剂三
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液体 18 mL×1 瓶
|
2-8℃保存
|
试剂四
|
液体 3 mL×1 瓶
|
2-8℃保存
|
粉剂一
|
粉剂×1 瓶
|
2-8℃保存
|
标准品
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粉剂×1 支
|
2-8℃保存
|
溶液的配制:
1、提取液配制:临用前根据样本量按照提取液一:提取液二=1mL : 1 mL进行配制,现配现用,请勿一次性全部混合。
2、若试剂二有析出,可置于37℃水浴加热至澄清透明后使用。
3、标准品:10mg还原型谷胱甘肽(GSH)。临用前加入1.3mL蒸馏水配制成25μmol/mL ,2-8℃保存4周。4 、0.125μmol/mL标准品配制:取50μL 25μmol/mL标准品,加入950μL蒸馏水,充分混匀,配制成 1.25μmol/mL的标准品;然后取100μL 1.25μmol/mL标准品,加入900μL蒸馏水,充分混匀,配制成0.125μmol/mL的标准品使用,现配现用。
产品说明:
巯基的存在使得蛋白质能够进行二硫键的形成,从而维持分子的稳定性和功能性。巯基还参与到氧化还原反应中,具有重要的生物学作用。在细胞内,巯基含量的变化与多种疾病的发生和进展密切相关,因此巯基也成为了生物医学领域中的重要研究对象。
本试剂盒测定的是蛋白质二硫键断裂产生的巯基和本身游离巯基的总和。
还原剂会使二硫键裂解生成巯基,巯基会发生亲核反应,即巯基与5,5’-二硫代-双-硝基苯甲酸(DTNB)反应,生成黄色化合物,在412nm处有吸收峰,据此可以计算蛋白质总巯基含量。
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)
注意:实验之前建议选择2-3个预期差异大的样本做预实验。如果样本吸光值不在测量范围内建议稀释或者增加样本量进行检测。
需自备的仪器和用品:
可见分光光度计、1mL玻璃比色皿、低温离心机、 研钵/匀浆器/细胞超声破碎仪、丙酮(AR)、蒸馏水。
操作步骤:
一、样本处理(可适当调整待测样本量)
1. 组织:按照质量(g):提取液体积(mL)为1:5~10的比例(建议称取约0.1g,加入1mL提取液)加入提取液,冰浴匀浆后于4℃ , 3000rpm 离心10min ,弃上清。在沉淀中加入2mL试剂一,搅匀后溶解沉淀,沉淀溶解液作为样本进行实验。
注:
1. 植物叶片等纤维含量较高的样本,溶解沉淀后4℃3000rpm离心3min ,取上清作为样本进行实验,;(2)加入试剂一后会有大量气泡产生,请缓慢加入,建议使用5mL的EP管)。
2. 细菌/细胞:按照细菌/细胞数量(106个):提取液体积(mL)为5~10:1的比例(建议5百万细菌/细胞加入1mL提取液),冰浴超声波破碎(功率200W,超声3秒,间隔10秒,总时间3min)后,于4℃ , 3000rpm离心10min,弃上清。在沉淀中加入2mL试剂一,搅匀后溶解沉淀,沉淀溶解液作为样本进行实验。
注:
1. 若沉淀溶解 不完全,可4℃3000rpm离心3min ,取上清作为样本进行实验;
2. 加入试剂一后会有大量气泡产生,请缓 慢加入,建议使用5mL的EP管)。
3. 血清/血浆、牛奶等液体:取100μL液体样本加入0.9mL丙酮,4℃ , 3000rpm离心10min ,弃上清。在沉淀中加入2mL试剂一,搅匀后溶解沉淀,沉淀溶解液作为样本进行实验。
注:若测定数值偏小,可改变样本与丙酮的比例,如取0.2mL液体样本加入0.8mL丙酮或0.3mL液体样本加入0.7mL丙酮,注意同步修改计算公式)。