计算溶液所需的质量、体积或浓度。
提供 ≥98%(HPLC) 纯度,适用于对基线干扰要求严格的色谱和分析工作流程。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
S Tag Peptide (S标签多肽),也称S-tag Peptide,是一种常用于通过竞争性结合Anti-S Tag抗体,从而在免疫沉淀时洗脱与Anti-S Tag抗体结合的S Tag融合表达蛋白的多肽。本产品不仅可以用于Anti-S Tag Magnetic Beads (Anti-S Tag磁珠)免疫沉淀时S Tag融合表达蛋白的洗脱,也可以用于常规的Anti-S Tag抗体免疫沉淀时S Tag融合表达蛋白的洗脱。本产品竞争性洗脱S Tag融合表达蛋白后,洗脱后的样品中不含Anti-S Tag抗体的轻链和重链,可以有效解决免疫沉淀后Western检测时轻链和重链的干扰问题。普通的免疫沉淀,通常是通过加入上样缓冲液煮沸从而使S Tag融合表达蛋白进入到溶液中,但此时Anti-S Tag抗体的轻链和重链也会进入到溶液中,并很可能会干扰后续的Western检测。S Tag Peptide (S Tag多肽)是合成的多肽,其氨基酸序列为Lys-Glu-Thr-Ala-Ala-Ala-Lys-Phe-Glu-Arg-Gln-His-Met-Asp-Ser (KETAAAKFERQHMDS)。S Tag Peptide (S Tag多肽)的主要参数如下:本产品5mg/ml包装产品配制在TBS中,可以直接使用。25mg为粉末装。S Tag Peptide (S Tag多肽)常用储存液浓度为5-25mg/ml,常用使用浓度为100-400μg/ml。
注意事项:
由于多肽的每次冻融均会引起部分失活,所以溶液装或粉末装配制成溶液后须分装后于-20℃或更低温度冻存,以避免反复冻融。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.对于粉末装,由于微量的多肽在冻干过程中沉积在管内,形成很薄或不可见的多肽层,所以在打开管盖前,建议在离心机中约8,000-12,000×g离心10-30秒,使附着在管盖或管壁上的多肽聚集于管底。2.对于粉末装,使用TBS进行溶解,一般用移液枪的枪头轻吹几下或者轻轻摇晃瓶子,即可使多肽完全溶解。请勿用vortex剧烈振荡,以免多肽变性而失活;溶液装可直接用于后续配制。3.免疫沉淀样品的S Tag Peptide竞争洗脱法:本方法为非变性法,洗脱效率高,且洗脱后的样品中不含Anti-S Tag抗体的轻链和重链。a.S Tag多肽洗脱液的配制:根据样品量确定洗脱液的用量,取一定量S Tag多肽储存液用TBS溶液进一步稀释S Tag多肽溶液至150μg/ml。b.每个免疫沉淀样品加入100μl S Tag多肽洗脱液(150μg/ml),冰浴并摇晃孵育30分钟-2小时,为了提高洗脱效率,可延长孵育时间或重复洗脱。S Tag多肽洗脱液体积一般为凝胶悬液或磁珠悬液的5倍。c.4℃,6000×g离心30秒,小心转移上清至新管中,上清即为洗脱的S Tag融合蛋白。注意不要吸到凝胶。4.洗脱的S Tag融合蛋白置于4℃待用,或者-20℃长期保存。The S Tag Peptide is commonly used for competitive elution of S tag fusion proteins bound to S Tag antibodies. It can be used to elute S tag fusion proteins from Anti-S Tag Magnetic Beads or conventional S Tag antibodies during immunoprecipitation.This product competitively elutes the S Tag fusion protein, and the eluted sample does not contain the light and heavy chains of S Tag antibodies, thus effectively eliminating the interference of antibodies with subsequent Western blot analysis.The main parameters of S Tag Peptide are as follows
Precautions:
The solution of this product should be aliquoted and stored at -20℃ or lower temperatures to avoid repeated freeze-thaw cycles.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. For 25mg provided in powder form, centrifuge at 8,000-12,000×g for 10-30 seconds before opening to collect the powder at the bottom of the tube. Dissolve it in TBS by gently pipetting or shaking. Do not vortex vigorously to avoid denaturation and inactivation of the peptide.2. P9816-1mg and P9816-5mg can be directly used for preparing elution buffer. 3. Competitive elution with the S Tag Peptide: This elution method is non-denaturing, with high elution efficiency, and the eluted sample does not contain the light and heavy chains of S Tag antibodies.a. Preparation of S tag elution buffer: Based on the required amount of elution buffer, dilute an appropriate amount of S Tag Peptide stock solution with TBS to a final concentration of 150μg/ml.b. Add 100μl of S tag elution buffer (150μg/ml) to each immunoprecipitated sample, and incubate on ice or at 4℃ for 30 minutes to 2 hours with shaking. In order to improve the elution efficiency, the incubation time can be extended or the elution can be repeated. The volume of the S tag elution buffer is generally 5 times that of the bead or gel suspension.c. Centrifuge at 6000×g for 30 seconds at 4℃, and carefully transfer the supernatant to a new tube. The supernatant contains the S tag fusion protein and its protein complexes. Be careful not to touch the resin when taking the supernatnant. 4. Store the eluted S tag fusion protein at 4℃ for immediate use, or at -20℃ for long-term storage.