计算溶液所需的质量、体积或浓度。
SDS-PAGE蛋白上样缓冲液 (6×) 是以溴酚蓝为染料,6倍浓缩的SDS-PAGE凝胶电泳上样缓冲液,具有沉淀蛋白样品进入凝胶孔和指示蛋白样品在凝胶中运动位置的作用。可使蛋白质分子的链内二硫键和链间二硫键断裂,各蛋白亚单位彼此分离以及蛋白变性的作用,适用于还原型SDS-PAGE的蛋白样品制备和上样。本产品使用了无气味、水溶性更稳定、还原能力相近的还原剂替代了有气味的二硫苏糖醇(DTT)或巯基乙醇(2-Mercaptoethanol),从而可以确保本SDS-PAGE蛋白上样缓冲液在正常使用或加热时都不会有异味,与常规SDS-PAGE蛋白上样缓冲液的使用效果一致,未观察到有任何显著差异。
使用说明:
1、试剂准备
1.1 在室温或不超过37°C的水浴中溶解 SDS-PAGE蛋白上样缓冲液 (6×) 。水浴溶解后立即室温存放,尽量避免长时间置于水浴中。
1.2 将 SDS-PAGE蛋白上样缓冲液 (6×) 与蛋白样品按照1:5的比例混匀;
2、蛋白样品处理
2.1 将蛋白样品置于沸水浴中加热5-10分钟;
2.2 待蛋白样品充分变性后冷却至室温,以小于3000rpm的条件离心30秒;
3、蛋白上样及电泳
3.1 离心后,以微量进样器取适量上清,直接加入SDS-PAGE凝胶加样孔内;
3.2 进行常规电泳,通常染料到达距离凝胶底端0.5cm-1cm处即可停止电泳;
注意事项:
1、加样前在室温或37°C水浴数分钟解冻后轻轻摇匀,以确保溶液混合均匀。
2、本产品仅用于科研,不能用于人体实验或人体治疗。
3、本产品用于蛋白变性时,建议95°C水浴,温度过高(如100°C)或时间过长(如超过15分钟),有可能会导致蛋白降解。
SDS-PAGE Protein Sample Loading Buffer (6×) uses bromophenol blue as the tracking dye. It facilitates loading protein samples into gel wells and monitors the migration front of samples during electrophoresis. It breaks intra‑ and inter‑chain disulfide bonds of proteins, separates protein subunits and denatures proteins. It is suitable for sample preparation and loading for reducing SDS‑PAGE. This product adopts an odorless, water‑soluble, stable reducing agent with comparable reducing capacity to replace malodorous dithiothreitol (DTT) or 2‑mercaptoethanol. Therefore, no unpleasant odor is generated during normal handling or heating. Its performance is consistent with conventional SDS‑PAGE loading buffers, with no significant differences observed.
Instructions for Use:
1. Reagent Preparation
1.1 Thaw SDS-PAGE Protein Sample Loading Buffer (6×) at room temperature or in a water bath not exceeding 37°C. Store at room temperature immediately after thawing; avoid prolonged incubation in the water bath.
1.2 Mix SDS-PAGE Protein Sample Loading Buffer (6×) with protein sample at a ratio of 1:5.
2. Protein Sample Treatment
2.1 Heat the protein sample in a boiling water bath for 5–10 minutes.
2.2 After complete denaturation, cool the sample to room temperature and centrifuge at less than 3000 rpm for 30 seconds.
3. Loading and Electrophoresis
3.1 Pipette an appropriate volume of the supernatant after centrifugation and load directly into the wells of the SDS‑PAGE gel.
3.2 Run standard electrophoresis. Stop the run when the dye front reaches 0.5–1 cm from the bottom edge of the gel.
Precautions:
1. Thaw the buffer at room temperature or in a 37°C water bath for several minutes and mix gently before use to ensure homogeneity.
2. For research use only. Not for human experiments or therapeutic applications.
3. A 95°C water bath is recommended for protein denaturation. Excessively high temperature (e.g., 100°C) or overlong incubation (more than 15 minutes) may cause protein degradation.
| pH | 6.5-7.5 |
|---|
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | S750311 |
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