计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
本产品能够快速、温和、高效地裂解哺乳动物细胞，有效提取细胞浆和细胞核蛋 白。该试剂使用温和配方保证所提取蛋白保持生物学活性并可应用于多种蛋白分析试 验，如报告基因和酶活性测定、免疫检测、蛋白纯化等。提取后蛋白可采用BCA法进 行蛋白定量分析。试剂盒中带有蛋白酶抑制剂混合物，可有效避免蛋白提取过程中蛋 白的降解。
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注意事项
1．本产品可有效裂解细胞培养板培养的贴壁细胞（无需刮取）以及离心收集的悬浮细胞， 较反复冻融或超声法具有更高提取效率。但对于组织蛋白的抽提，建议使用组织蛋白抽 提试剂盒（CW0891）。
2．表1中所列的是贴壁细胞蛋白提取的最佳使用量，先收集细胞可以减少试剂用量从而获 得更高的蛋白浓度。
3．也可以根据细胞数量估计抽提试剂的使用量。如2×106 个Hela细胞约重20 mg，需要加入 200 μl抽提试剂。
4．通过本产品提取的蛋白，可通过BCA法进行蛋白定量分析。
操作步骤
● 贴壁细胞蛋白抽提
1．请在蛋白抽提前取出实验所需Mammalian Protein Extraction Reagent进行预冷。
2．小心倾去贴壁细胞的培养液，使用PBS漂洗细胞。
3．加入适量Mammalian Protein Extraction Reagent（抽提蛋白前2-3分钟按照1:99比 例加入Protease Inhibitor Cocktail），在冰上用枪头吹打贴壁细胞，将裂解液转移至 离心管中，冰上孵育20分钟，让细胞充分裂解（试剂使用量请参考附表1，冰上放 置时间应根据细胞类型不同进行调整）。
4．14,000×g离心5-10分钟。
5．转移上清液至新管中，用于进一步分析。
● 悬浮细胞蛋白提取
1．请在蛋白抽提前取出实验所需Mammalian Protein Extraction Reagent进行预冷。
2．将悬浮细胞2,500×g，离心10分钟，弃去上清。使用PBS漂洗细胞。2,500×g，离心 10分钟，弃去上清。
3．加入适量Mammalian Protein Extraction Reagent，抽提蛋白前2-3分钟按照1:99比 例加入Protease Inhibitor Cocktail，即1×工作液。
4．每100 mg细胞加入至少1 ml 1×工作液。如提取的样本量较大，可首先使用少量1×工 作液重悬细胞，然后加入剩余工作液。
5．吹打均匀后，冰上放置20分钟，让细胞充分裂解（冰上放置时间应根据细胞类型不 同进行调整）。
6．14,000×g离心15分钟。
7．转移上清至新管中，进行下一步分析。
表1. 抽提试剂使用量推荐表
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This product can rapidly, gently, and efficiently lyse mammalian cells, effectively extracting cytoplasmic and nuclear proteins. This reagent uses a mild formula to ensure that the extracted protein maintains biological activity and can be applied to various protein analysis experiments, such as reporter gene and enzyme activity determination, immune detection, protein purification, etc. The extracted protein can be quantitatively analyzed using the BCA method. The reagent kit contains a mixture of protease inhibitors, which can effectively prevent protein degradation during the protein extraction process.
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precautions
1. This product can effectively lyse adherent cells cultured on cell culture plates (without scraping) and suspended cells collected by centrifugation, with higher extraction efficiency than repeated freeze-thaw or ultrasound methods. But for the extraction of tissue proteins, it is recommended to use the tissue protein extraction kit (CW0891).
The optimal dosage for protein extraction from adherent cells is listed in Table 1. Collecting cells first can reduce the amount of reagents used to obtain higher protein concentrations.
3. The amount of extraction reagents used can also be estimated based on the number of cells. If 2 × 106 Hela cells weigh about 20 mg, 200 need to be added μ Extract reagents.
4. The protein extracted from this product can be quantitatively analyzed using the BCA method.
Operation steps
● Protein extraction from adherent cells
1. Please remove the required Mammalian Protein Extraction Agent for pre cooling before protein extraction.
2. Carefully pour out the culture medium of adherent cells and rinse the cells with PBS.
3. Add an appropriate amount of Mammalian Protein Extraction Reagent (add Protein Inhibitor Cocktail in a 1:99 ratio 2-3 minutes before protein extraction), blow adherent cells on ice with a gun tip, transfer the lysate to a centrifuge tube, incubate on ice for 20 minutes, and allow the cells to fully lyse (please refer to Appendix 1 for the amount of reagent used, and the time for placing on ice should be adjusted according to different cell types).
4. Centrifuge at 14000 × g for 5-10 minutes.
5. Transfer the supernatant to a new tube for further analysis.
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Suspension cell protein extraction
1. Please remove the required Mammalian Protein Extraction Agent for pre cooling before protein extraction.
2. Suspend 2500 × g of cells, centrifuge for 10 minutes, and discard the supernatant. Rinse cells with PBS. 2500 × g, centrifuge for 10 minutes, discard the supernatant.
3. Add an appropriate amount of Mammalian Protein Extraction Agent, and 2-3 minutes before protein extraction, add Protein Inhibitor Cocktail in a ratio of 1:99, which is 1 x working solution.
4. Add at least 1 ml of 1x working solution to every 100 mg of cells. If the extracted sample size is large, a small amount of 1x working solution can be used to resuspend the cells first, and then the remaining working solution can be added.
5. After blowing evenly, place it on ice for 20 minutes to allow the cells to fully lyse (the time for placing it on ice should be adjusted according to different cell types).
6. Centrifuge at 14000 × g for 15 minutes.
7. Transfer the supernatant to a new tube for further analysis.
Table 1. Recommended usage of extraction reagents
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Table 2. Common Problems and Solutions
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规格
英文名称
Mammalian Protein Extraction Kit
储存与运输
储存条件
-20°C储存
运输条件
超低温运输
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
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溶液计算器
常见问题
本产品应如何储存？
请于?20 °C条件下保存。需冷冻保存以维持规定的有效期。
本产品如何运输？
本产品采用保温箱配冰袋运输。收货后请立即拆包，并按上述储存条件保存。
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