双色SDS-PAGE蛋白上样缓冲液

D752143-2ml

2ml

6X, 无气味

阿拉丁生产的双色SDS-PAGE蛋白上样缓冲液(6X, 无气味) (Dual Color SDS-PAGE Protein Sample Loading Buffer, 6X, Odorless)，是一种经过改良的更加安全健康的无气味的使用蓝色和红色染料、6倍浓缩的蛋白上样缓冲液。 本产品更加安全健康。本产品使用了无异味、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，使蛋白上样操作更加安全健康。 除了没有气味外和染料不同外，本产品和常规的蛋白上样缓冲液的使用效果一致。本产品可以用于常规的SDS-PAGE蛋白样品的上样。除了没有异味和染料不同外，本产品与常规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。 用于常规的SDS-PAGE电泳时，本产品中蓝色染料的迁移率与溴酚蓝一致，红色染料的迁移速度和蓝色染料相近。Tris-Gly电泳体系中，在SDS-PAGE的胶浓度低于15%时，红色染料的迁移速度微快于蓝色染料，胶浓度高于15%时，红色染料的迁移速度微慢于蓝色染料。 本产品可以用于转膜后的泳道示踪：本双色SDS-PAGE蛋白上样缓冲液(6X, 无气味)中含特殊的红色染料，该红色染料可随蛋白转印到PVDF或NC等印迹膜上，从而可以用于泳道位置的指示，便于剪膜等操作。双色类SDS-PAGE蛋白上样缓冲液产品的电泳及转膜效果示意图参见图1。 图1. 双色类SDS-PAGE蛋白上样缓冲液产品用于UltraBio™ Plus PAGE预制胶(Tris-Gly, 12%, 10孔) 的电泳后的凝胶图(Gel after electrophoresis)、转膜后PVDF膜图(Blot)和转膜后凝胶图(Gel after transfer)。 注意事项 ： 本产品用于蛋白变性时，建议95℃水浴或PCR仪加热5分钟，温度过高(如100℃)或时间过长(如超过15分钟)，有可能会导致蛋白降解或上样缓冲液中指示剂的颜色异常。 本产品不含剧毒的巯基乙醇和有刺激性气味的DTT，但还原效果一致，对于蛋白样品的处理效果和电泳效果一致。 本产品必须完全溶解后再使用。 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。 为了您的安全和健康，请穿实验服并戴一次性手套操作。 使用说明 ： 1.在室温或不超过37℃的水浴中溶解双色SDS-PAGE蛋白上样缓冲液(6X, 无气味)。水浴溶解后立即室温存放，尽量避免长时间置于水浴中。使用完毕后置于-20℃保存。2.按照每5微升蛋白样品加入1微升双色SDS-PAGE蛋白上样缓冲液(6X, 无气味)的比例，混合蛋白样品和双色蛋白上样缓冲液(6X, 无气味)。注：本产品稀释至1X后也可以直接用于细胞或组织样品的裂解。3.95℃水浴加热5-10分钟，以充分变性蛋白。说明：加热前通常会发现蛋白样品内有粘稠的半透明状物体，通常在本上样缓冲液内95度水浴加热8-10分钟后可以使该粘稠的半透明状物体消失，以便于后续的上样操作。注意：请务必95℃加热，温度过高(如100℃)或时间过长(如超过15分钟)，有可能会导致蛋白降解或上样缓冲液中指示剂的颜色异常。注意：如果起始时细胞或组织的用量较大，基因组DNA含量较高，加热5-10分钟后有可能仍然比较粘稠或者有粘稠状的半透明物体。此时需要再加热5-10分钟或者加入适量1X的蛋白上样缓冲液后再加热3-5分钟。充分加热后一方面可以使结合在基因组DNA上的蛋白充分释放，同时会导致基因组DNA的部分断裂从而使粘稠感消失，这样就不会影响后续的上样操作了。适当超声或使用1ml注射器反复抽吸也可以打断基因组DNA从而使粘稠感消失。4.冷却到室温后，直接上样到SDS-PAGE胶加样孔内即可。5.通常电泳至红色或蓝色染料到达胶的底端处附近即可停止电泳。 Aladdin's Dual Color SDS-PAGE Protein Sample Loading Buffer (6X, Odorless) is an improved protein loading buffer with red and blue dyes, which can be used for the preparation and loading of protein samples onto a gel for SDS-PAGE analysis.This product is safe to use. Instead of odoriferous DTT or 2-Mercaptoethanol, this product employs an odorless and more stable reducing agent with similar reducing capability.In conventional SDS-PAGE, the blue dye migrates exactly the same as bromophenol blue, and the red dye migrates similarly to the blue dye. In Tris-Gly electrophoresis system, the red dye migrates slightly faster than the blue dye in SDS-PAGE gel lower than 15%. While in SDS-PAGE higher than 15%, the red dye migrates slightly slower than the blue dye.This product can be used for lane tracing after membrane transfer. It contains a special red dye which can be transferred to a PVDF or NC membrane to indicate the position of the lane, facilitating operations such as film cutting. Please refer to Figure 1 for the electrophoresis and the transfer results of this product.Figure 1. Aladdin's Dual Color SDS-PAGE Protein Sample Loading Buffer (6X, Odorless) in the UltraBio™ Plus PAGE Precast Gel (Tris-Gly, 12%, 10 wells) after electrophoresis and transfer, as well as in the PVDF membrane after transfer (Blot). Precautions ： For denaturing PAGE analysis, protein samples mixed with this product is recommended to be heated at 95℃ for 5 minutes. Too high a temperature (e.g., 100℃) or too long a time (e.g., more than 15 minutes) may result in protein degradation or abnormal color change of the indicator.This product must be completely dissolved and mixed well before use.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation. Instructions for Use ： 1. Thaw this product at room temperature or in a water bath no more than 37℃. Place at room temperature immediately once dissolved and store at -20℃ after use.2. Add 1μl of this product per 5μl of protein sample, and mix well. Note: After dilution to 1X, this product can also be used for sample lysis directly.3. Heat at 95℃ for 5-10 minutes to fully denature proteins.Note: If the starting material is large and the genomic DNA content is high, the mixture may be very sticky after boiling. In this case, heat for another 5-10 minutes directly or after addition of more 1X sample loading buffer. The thorough heating will release proteins bound to genomic DNA and break genomic DNA so that the mixture will be not too sticky for sample loading. Appropriate ultrasound or repeated aspiration with a 1ml pipette can also help.4. After cooling to room temperature, centrifuge briefly at room temperature, then load the supernatant onto a SDS-PAGE gel.5. Stop electrophoresis when the red or blue dye reaches the bottom of the gel.