SDS-PAGE上层胶预混液

阿拉丁生产的SDS-PAGE上层胶预混液(SDS-PAGE Stacking Gel Master Mix)提供了简单而又超快速地配制SDS-PAGE凝胶(即SDS聚丙烯酰胺凝胶)上层胶所需的主要试剂，包括Tris-HCl (pH6.8)、Acr-Bis(29:1)、SDS等。阿拉丁生产的SDS-PAGE上层胶预混液为最常用的5%浓度上层胶，如您需要配制特殊浓度的上层胶，可以考虑单独购买ST003 Acr-Bis(29:1)、ST628 10%SDS和ST768 Tris-HCl (pH6.8)等进行特殊浓度SDS-PAGE凝胶上层胶的配制。本上层胶预混液已经把SDS-PAGE凝胶上层胶配制所需的Tris-HCl (pH6.8)、Acr-Bis(29:1)、SDS等预混合好，使用前仅需加入适量凝胶聚合催化剂(过硫酸铵或其替代物，如ST005 Ammonium persulfate substitute)和ST728 TEMED即可简单快速地完成上层胶(分离胶)的配制。本上层胶预混液可以直接配合阿拉丁不同浓度的SDS-PAGE下层胶预混液使用，轻松超快速完成各种不同浓度SDS-PAGE凝胶的配制。本产品的250ml包装约可配制50-100块常规大小的SDS-PAGE凝胶的上层胶，500ml包装约可配制100-200块常规大小的SDS-PAGE凝胶的上层胶。具体可以配制的凝胶数量和凝胶的厚薄以及凝胶的大小有关。 注意事项 ： 进行上层胶配制时，需要另外准备过硫酸铵或其替代物以及TEMED 。必须在下层胶配制完成后才能进行上层胶的配制。本产品含有Acr-Bis，对人体有毒，操作时请特别小心，并注意有效防护以避免直接接触人体或吸入体内。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.根据目的蛋白的分子量大小选择合适浓度的SDS-PAGE下层胶(即分离胶)。不同浓度的SDS-PAGE分离胶的最佳分离范围如下：SDS-PAGE分离胶浓度最佳分离范围6%胶50-150kD8%胶30-90kD10%胶20-80kD12%胶12-60kD15%胶10-40kD2.称取适量过硫酸铵或其替代物，如ST005，用双蒸水或其它高纯度的水配制10%溶液。过硫酸铵或其替代物配成溶液后容易失效，需注意存放及时间限制。3.参考下表，在下层胶凝固后，在SDS-PAGE上层胶预混液中，按照1%的比例加入相应量的10%过硫酸铵或其替代物溶液，按照0.1%的比例加入相应量的TEMED。例如10mlSDS-PAGE上层胶预混液中，加入100μl10%过硫酸铵或其替代物溶液和10μl TEMED，适当混匀。去除下层胶上面覆盖的液体，尽量去干净，然后倒入添加了10%过硫酸铵或其替代物溶液和TEMED的SDS-PAGE上层胶预混液，插入梳子待凝固。上层胶凝固后，则表明制胶步骤结束，可以准备进行后续的电泳了。通常10-30分钟内胶会凝固。注：具体的凝固时间和温度及光照有关，说明书中10%过硫酸铵或其替代物和TEMED的正常推荐用量是室温为25℃时的推荐用量。为达到与25℃时相近的凝固时间，当室温低于25℃时，可以适当同时加大10%过硫酸铵或其替代物和TEMED的用量，例如20℃时建议用量是正常推荐用量的1.5倍，15℃时建议用量是正常推荐用量的2倍。成分配制不同体积SDS-PAGE上层胶所需各成分的体积(毫升)SDS-PAGE上层胶预混液234681010%过硫酸铵或其替代物0.020.030.040.060.080.1TEMED0.0020.0030.0040.0060.0080.014.配制好的凝胶如果当天不能使用，可以在4℃保存1-2天后使用。 Aladdin's SDS-PAGE Stacking Gel Master Mix contains almost all reagents required for the preparation of SDS-PAGE stacking gels, except for polymerization catalysts such as ammonium persulfate and TEMED.This product can be used to prepare the most commonly used 5% stacking gels. If other concentrations are desired, we recommend purchasing Acr-Bis (29 Precautions ： Ammonium persulfate or its substitute (, ST005) and TEMED are required but not provided in this product.The stacking gel must be prepared after the resolving gel is polymerized.This product contains Acr-Bis which is potentially neurotoxic. Please take effective measures to avoid direct contact with the human body 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 Range6%50-150kD8%30-90kD10 -80kD12%12-60kD15%10-40kD2.Prepare a 10% ammonium persulfate solution with ddH2O or other high-purity water. Solution of ammonium persulfate or its substitute is prone to failure and should be prepared freshly or kept frozen for multiple uses.3.After the resolving gel is fully polymerized, prepare the stacking gel according to the table below. For example, add 30μl of 10% ammonium persulfate and 3μl of TEMED into 3ml of SDS-PAGE Stacking Gel Master Mix, mix well, and immediately pour onto the top of the resolving gel without sealing liquid. Insert the comb and leave at room temperature (~25℃) until fully solidified (usually within 10-30 minutes).Note: The amount of polymerization catalyst in the table below is recommended for polymerization at 25℃. It can be adjusted appropriately according to the temperature. For example, when the room temperature is lower than 20℃, add more TEMED and 10% ammonium persulfate appropriately to promote gel solidification. ReagentsVolume of each component (ml) required for different volumes of SDS-PAGE resolving gelSDS-PAGE Stacking Gel Master Mix234681010% ammonium persulfate or its substitute0.020.030.040.060.080.1TEMED 0.0020.0040.0060.0080.0120.024.If the prepared gel is not to be used on the same day, it can be stored at 4℃ for 1-2 days.

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