非变性蛋白上样缓冲液 (5×, 含TCEP) 是一种用于蛋白质电泳分析的缓冲液，其主要特点是不含变性剂（如SDS）和还原剂（如DTT），但含有还原剂TCEP（Tris(2-carboxylethyl)phosphine）。该缓冲液不含变性剂，因此不会破坏蛋白质的二级和三级结构，使得蛋白质在电泳过程中保持其天然状态。TCEP可替代DTT或巯基乙醇还原二硫键，但无DTT或巯基乙醇等还原剂的有害的刺鼻气味。
这种缓冲液主要用于保持蛋白质的天然构象，以便在非变性条件下进行电泳分析。本品可以用于常规的非变性聚丙烯酰胺凝胶（PAGE）蛋白样品的上样。
该试剂仅用于科研领域，不适用于临床诊断或其他用途。
操作步骤（仅供参考）：
1、溶解缓冲液：在室温或不超过37℃的水浴中溶解非变性蛋白上样缓冲液 (5×，含TCEP)。溶解后应立即室温存放，避免长时间置于水浴中。
2、样品准备：按照每4 μL蛋白样品加入1 μL 非变性蛋白上样缓冲液 (5×，含TCEP)的比例，混合蛋白样品和缓冲液。
3、加热处理：将混合后的蛋白样品置于95℃中加热3-5分钟，以确保蛋白质充分变性。
4、冷却和离心：待蛋白样品充分变性后冷却至室温，并以小于3000 rpm的条件离心30秒，去除可能产生的气泡和沉淀。
5、上样：使用微量进样器取适量上清，直接加入PAGE凝胶加样孔内。
6、电泳：进行常规电泳，通常当蓝色染料到达距离凝胶底端0.5-1 cm处即可停止电泳。
注意事项：
1、确保非变性蛋白上样缓冲液 (5×，含TCEP) 完全溶解后再使用。
2、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。
3、为了您的安全和健康，请穿实验服并戴一次性手套操作。
4、试剂开封后请尽快使用，以防影响后续实验效果。
Native-PAGE Loading Buffer (5×, with TCEP) is a buffer used for protein electrophoresis analysis. Its main feature is that it does not contain denaturants (such as SDS) or reducing agents (such as DTT), but contains the reducing agent TCEP (Tris(2-carboxyethyl)phosphine). Since this buffer does not contain denaturants, it does not disrupt the secondary and tertiary structures of proteins, allowing proteins to maintain their native state during electrophoresis. TCEP can replace DTT or mercaptoethanol to reduce disulfide bonds, but it does not have the harmful pungent odor of reducing agents such as DTT or mercaptoethanol.
This buffer is mainly used to maintain the native conformation of proteins so that electrophoresis analysis can be performed under non-denaturing conditions. This product can be used for loading conventional non-denaturing polyacrylamide gel (PAGE) protein samples.
This reagent is for research use only and is not suitable for clinical diagnosis or other purposes.
Procedure (for reference only):
1.Dissolve the buffer: Dissolve Non-denaturing Protein Loading Buffer (5×, containing TCEP) at room temperature or in a water bath not exceeding 37 ℃. After dissolution, store it at room temperature immediately and avoid leaving it in the water bath for a long time.
2.Sample preparation: Mix the protein sample and buffer at a ratio of 1 μL of Non-denaturing Protein Loading Buffer (5×, containing TCEP) per 4 μL of protein sample.
3.Heat treatment: Place the mixed protein sample at 95 ℃ for 3–5 minutes to ensure that the protein is fully denatured.
4.Cooling and centrifugation: After the protein sample is fully denatured, cool it to room temperature and centrifuge at less than 3000 rpm for 30 seconds to remove possible bubbles and precipitates.
5.Loading: Use a microsyringe to take an appropriate amount of supernatant and directly add it into the wells of the PAGE gel.
6.Electrophoresis: Perform routine electrophoresis. Usually, electrophoresis can be stopped when the blue dye reaches 0.5–1 cm from the bottom of the gel.
Precautions:
1.Ensure that Non-denaturing Protein Loading Buffer (5×, containing TCEP) is completely dissolved before use.
2.This product is limited to scientific research use by professionals. It must not be used for clinical diagnosis or treatment, must not be used for food or drugs, and must not be stored in ordinary residential buildings.
3.For your safety and health, please wear a laboratory coat and disposable gloves during operation.
4.After opening the reagent, please use it as soon as possible to prevent affecting subsequent experimental results.
危险品化学品经营许可证（带存储）