SDS-PAGE One-step Resolving Gel Master Mix (C007B-566108)

Aladdin's SDS-PAGE One-step Resolving Gel Master Mix contains almost all reagents required for the simple and ultra-fast preparation of DS-PAGE resolving gels at concentrations of 6%, 8%, 10%, 12%, and 15%, except for polymerization catalysts such as ammonium persulfate (APS) and TEMED. It contains Tris-HCl (pH 8.8), Acr-Bis (29Precautions：Ammonium Persulfate or its substitute (, ST005) and TEMED are required but not provided in this product.This product contains Acr-Bis which is potentially neurotoxic. Please take effective measures to avoid direct contact or inhalation.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.According to the size of a target protein, decide an appropriate concentration of SDS-PAGE resolving gel. Please refer to the table below. SDS-PAGE Resolving Gel Concentration Optimal Separation Range 6% 50-150kDa 8% 30-90kDa 10% 20-80kDa 12% 12-60kDa 15% 10-40kDa 2.Prepare a 10% APS solution with ddH2O or other high-purity water (, ST872). Solution of APS or its substitute is prone to failure and should be prepared freshly or kept frozen for multiple uses. 3.Prepare the required volumes of resolving gel and stacking gel. For a gel plate with a width of 10cm, please refer to the table below to prepare a resolving gel with a height of 6cm and a stacking gel with a height of 1.5cm. Gel thickness Volume of the resolving gel Volume of the stacking gel 0.75mm 4.0ml 1.0ml 1.0mm 5.4ml 1.5ml 1.5mm 8.0ml 2.0ml Note: Do not cast too much resolving gel. Otherwise, there will be no sufficient space for the stacking gel.4.Prepare the resolving gel according to the table below. For example, add 100μl of 10% ammonium persulfate and 4μl of TEMED into 10ml of 10% SDS-PAGE Resolving Gel Master Mix, mix well, and immediately pour into the gel cassette. Afterwards, directly and gently add the stacking gel onto the the top of the resolving gel. Insert the comb and leave at room temperature (~25℃) until fully solidified. The stacking gel can be prepared using ’s SDS-PAGE One-step Red or Blue Stacking Gel Master Mix . Alternatively, before preparing the stacking gel, overlay the resolving gel with the top layer of water-saturated butanol, isopropanol, 0.1% SDS or distilled water to allow for gel polymerization for 10-30 minutes.Note 1: It is recommended to prepare a 4% stacking gel to achieve better layering effect. Other concentrations of stacking gel also can be used as appropriate.Note 2: When adding the stacking gel, it is important to gently and slowly add it with a 1ml pipette while moving it left and right, instead of pouring it in one spot. This is to prevent the stacking gel from rushing into the resolving gel and disrupting the smoothness of the gel interface.Note 3: When directly adding the stacking gel for one-step method, the smoothness of the gel interface may be slightly worse compared to the liquid sealing method, but it does not affect the electrophoresis results.Note 4: The polymeriztion time varies depending on temperature and light conditions. The recommended amount of 10% APS and TEMED is for use at room temperature (25℃). At temperature lower than 25℃, it may be necessary to increase the amount of 10% APS and TEMED. For example, we recommend using 1.5 times the normal amount at 20℃, and 2 times the normal amount at 15℃. Reagents Volume of each component (ml) required for different volumes of SDS-PAGE resolving gel 10% SDS-PAGE One-step Resolving Gel Master Mix 5 10 15 20 30 50 10% APS or its substitute 0.05 0.1 0.15 0.2 0.3 0.5 TEMED 0.002 0.004 0.006 0.008 0.012 0.02 5.If the prepared gel is not to be used on the same day, it can be stored at 4℃ for 1-2 days. However, we recommend using freshly prepared gels to get better resolution effect.6.For electrophoresis and Western blot procedures, please refer to https://www.aladdin-e.com.References：1.Laemmli UK. Nature. 1970. 227(5259):680-5.Related Products:Cat. 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