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Blot鉴定目标蛋白是否有His-Tag。若没有，需要重新构建载体，添加组氨酸标签，并确认标签正确表达。\nb、蛋白折叠导致组氨酸标签未完全暴露。\n解决措施：\n 在样品和平衡缓冲缓冲液中加1-2M尿素，这样蛋白结构相对松散，也许能吸附而蛋白不会变性。\n 在变性条件下纯",{"faq_id":250,"question":251,"answer_summary":252},"FAQ-20260930-1681db50","14.洗脱产物杂带多，是什么原因？","（1）杂蛋白吸附。由于组氨酸，色氨酸，半胱氨酸等是蛋白质中很常见基团，而蛋白折叠可能导致几个这样的氨基酸残基临近，这样也会使它们和磁珠的作用力增加。可以用不同浓度的咪唑进行梯度洗脱，在样品及平衡缓冲液中添加0.5%吐温或Triton X-100，或5~50%的甘油，可以避免因为疏水相互作用导致非特异吸附，或使用钴离子螯合磁珠（AnnbeadsTM IDA-Cobalt）。此外，要获得纯度较高的蛋白",{"faq_id":254,"question":255,"answer_summary":256},"FAQ-20260930-1896aa08","13.目标蛋白难洗脱","穿透组分中目标蛋白明显减少，而洗脱组分中目标蛋白很少，取少量磁珠加SDS-PAGE Loadding Buffer于95℃加热5~10min，离心跑电泳，如果发现较多蛋白残留。\n（1）洗脱调节太温和。提高洗脱液咪唑浓度至700mM还不能洗脱时，可以尝试降低洗脱液pH至4.0，但低pH会也导致金属离子脱落，磁珠需要再生后再使用。\n（2）蛋白吸附在磁珠上无法洗脱。对于非特异性疏水吸附的蛋白，可以增加N",{"faq_id":258,"question":259,"answer_summary":260},"FAQ-20260930-29059b78","5.磁珠黏壁现象是什么原因？如何解决？","由于疏水性蛋白黏附在磁珠上导致磁珠黏壁。建议缓冲液中加入0.05% tween-20。如果不能改善，不建议继续重复使用。",{"faq_id":262,"question":263,"answer_summary":264},"FAQ-20260930-44a77329","12.磁珠再生后，蛋白纯化效果变差，如何解决？","①再生过程的碱处理可以改善再生磁珠的蛋白纯化效果，参考说明书中磁珠再生部分步骤4。\n②重新挂金属离子后为清洗干净，需要增加洗涤步骤。残留的金属离子优先跟蛋白结合，影响蛋白与磁珠的结合。",{"faq_id":266,"question":267,"answer_summary":268},"FAQ-20260930-51c0b2eb","11.缓冲液中咪唑添加的意义？","结合缓冲液中低浓度的咪唑是为了减少杂蛋白的非特异性吸附；\n洗涤缓冲液中低浓度咪唑是为了吸取吸附能力较弱的杂蛋白；\n洗脱液中高浓度咪唑是为了洗脱目的蛋白。",{"faq_id":270,"question":271,"answer_summary":272},"FAQ-20260930-64b23044","6.磁珠适宜的pH值范围是多少？","磁珠适宜的pH值范围为pH3-11。",{"faq_id":274,"question":275,"answer_summary":276},"FAQ-20260930-788f7c54","4.若体系中含有木瓜蛋白酶，是否影响磁珠的使用？","木瓜蛋白酶是一种蛋白水解酶，目前对protein 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