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| 产品名称 | UltraBio™ His-tag Purification Resin | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| 别名 | UltraBio™ His-tag 纯化树脂 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 产品介绍 |
耐还原螯合型： 阿拉丁生产的UltraBio™ His-tag Purification Resin(耐还原螯合型)，即His标签蛋白纯化介质，俗称镍柱，是一种新型的可以兼容还原剂和螯合剂，并能简单、快速、高效并且高特异性地纯化His标签蛋白的纯化介质。
注意事项： Fast Flow, 耐还原螯合型：
1.1 介质清洗 方法一：取UltraBio™ His-tag Purification Resin介质，转移至G3砂芯漏斗中抽滤，加入去离子水搅拌、洗涤、抽干，重复洗涤抽干3次，最后抽干20 min。将介质用去离子水重悬，超声30 min去除气泡，超声过程中轻轻搅拌介质。 方法二：取 UltraBio™ His-tag Purification Resin介质，静止，待介质完全沉淀后，弃去上清液，加入与介质等体积或更多去离子水搅拌、清洗，静止，弃去上清液，重复清洗3次待用。 1.2 层析柱介质装填 1.2.1 检查仪器设备稳定性 取待装填层析柱，检查装柱器、层析柱上下适配器和柱管的完整性，包括胶圈是否缺失或损坏、连接软管是否通畅完整、接头状态、筛板是否丢失、筛网是否破损、玻璃柱是否有裂痕等，检查层析系统是否存在背景压力。 1.2.2 固定层析柱 确认层析柱和层析系统无误后，用去离子水清洗层析柱（如有需要可超声清洗），将层析柱下端适配器连接到层析系统，排出适配器内气泡，再连接到层析柱管下端，并保留约1cm高度的装柱液（防止加入填料时柱底部产生气泡），将装柱器连接到柱管上端，固定层 析柱垂直于桌面摆正。 1.3 恒流装柱 1) 装柱前将所有材料控制在相同温度（室温或4℃）。 2) 用玻璃棒搅拌将介质重悬，搅拌过程不可使用磁力搅拌器等剧烈搅拌方式，同时过程中 不可用玻璃棒敲打烧杯内壁，以防介质因外力受损。将重悬后的介质一次性倒入层析柱 中，倒入过程中使用玻璃棒轻靠层析柱内壁进行引流，防止倒入过程中产生气泡。 3) 将介质倒入层析柱后，快速连接装柱器上端，并打开层析系统泵，流速范围30-40cm/h，待介质胶面稳定。稳定后，取下装柱器，将层析柱适配器上端连接至层析系统，并排出内部气泡，插入层析柱中，在距离胶面上端1-2 cm处固定。 1.4 恒压装柱 将层析系统设置为0.1 MPa恒压装柱，时间1h，用 Marker笔记录最终胶面位置。恒压装柱结束后，将层析柱下端封死，断开层析柱上端与系统连接，轻松上端适配器密封胶圈，将上适配器下压至胶面下2mm处，拧紧密封圈，静止层析柱10 min，装柱结束。 1.5 柱效检测 方法一：丙酮检测方法
方法二：氯化钠检测方方法
2. 样品准备 1) 样品溶液需离心后，经0.22或0.45 μm滤膜过滤，去除细小颗粒物，防止堵塞层析柱和层析系统。 2) 样品缓冲液pH值和电导率值与平衡缓冲液保持一致或接近。 3. 平衡 1) 推荐平衡缓冲液：20 mM磷酸盐，0.5 M NaCl，pH值7.4；（优化：可加入低浓度 咪唑，避免杂蛋白结合，具体咪唑浓度根据实验结果确定，一般浓度范围5-20 mM）。 2) 流速：90-150 cm/h。 3) 用平衡缓冲液平衡层析柱5CV（通常3-5CV），当流出液的pH值和电导率与平衡缓冲液相同时，表明层析柱完全平衡。 4.上样 1) 将预处理好的样品泵入层析柱，流速90-150 cm/h。 2) 在足够的柱高或较低流速下，充分的保留时间，有助于目的蛋白与介质结合。 5.淋洗 上样结束后，继续用平衡液冲洗层析柱，去除因其他因素与介质结合的杂质，冲至紫外 平稳接近基线即可。 6. 洗脱 1) 推荐洗脱缓冲液 20 mM PB，0.5 M NaCl，500 mM 咪唑，pH 值7.2（为了提高目标蛋白的纯度，可采用含有不同浓度的咪唑缓冲液进行梯度洗脱）。 2) 其它洗脱方式 pH 洗脱：调节洗脱缓冲液pH值范围在2.5-5.0进行洗脱。如果蛋白质对低pH值敏感，建议在含有适当体积的中和液（1 M Tris-HCl，pH 9.0）的试管中收集pH洗脱液，以恢复中性pH值。 7. 在线清洗 当层析柱工作时反压明显增大，即表明介质需要进行在线清洗。清洗前，需要预先去除金属离子，并执行反向清洗操作。根据吸附蛋白的不同种类，建议按如下条件进行清洗。 1) 去除离子结合蛋白 1.5 M NaCl 缓冲液，清洗2CV。 2) 去除弱疏水结合蛋白、沉淀蛋白、脂蛋白 1 M NaOH 溶液，正/反向清洗1-2CV，直至杂质流出。 3) 去除强疏水结合蛋白、脂蛋白、脂质 70%乙醇或30%的异丙醇，反向清洗3-5CV。 清洗后，均用纯水清洗5-10CV。再根据实验计划进行结合缓冲液平衡或保存处理。 注：使用高浓度有机溶剂清洗液时，应逐步增加有机溶剂浓度，避免层析柱中产生气泡。 8. 保存 1) 未开封的介质可以储存在2-8 ℃环境中。 2) 使用过的介质需用20%乙醇进行置换，储存在4-8 ℃环境中。 耐变性剂型：
阿拉丁生产的UltraBio™ His-tag Purification Resin (耐变性剂型)，即
UltraBio™ His-tag Purification Resin (Denaturant- resistant)，是一种耐变性剂型的His标签蛋白纯化介质，俗称镍柱。本产品是一种新型的可以兼容尿素和盐酸胍等强蛋白变性剂，从而不仅可以在非变性条件下也可以在变性条件下简单、快速、高效并且高特异性地纯化His标签蛋白的纯化介质。
注意事项： Fast Flow,耐变性剂型： 阿拉丁生产的UltraBio™ His-tag Purification Resin (Fast Flow, 耐变性剂型)，即 阿拉丁生产的UltraBio™ His-tag Purification Resin (Fast Flow, Denaturant-resistant)，是一种能耐受高流速和高压力的耐变性剂型的His标签蛋白纯化介质，俗称镍柱。本产品是一种新型的能耐受高流速和高压力的可以兼容尿素和盐酸胍等强蛋白变性剂，从而不仅可以在非变性条件下也可以在变性条件下简单、快速、高效并且高特异性地纯化His标签蛋白的纯化介质。 通常带有6个组氨酸标签的重组蛋白(6xHis-tagged recombinant protein)或带有6个以上连续组氨酸标签的重组蛋白，被称为His标签蛋白。蛋白样品溶液通过 阿拉丁生产的UltraBio™ His-tag Purification Resin (Fast Flow, 耐变性剂型)时，重组蛋白His标签上的组氨酸残基能特异性地结合到 阿拉丁生产的UltraBio™ His-tag Purification Resin (Fast Flow, 耐变性剂型)中的镍离子上，其它蛋白则不能被结合。洗涤后，His标签重组蛋白可在非变性条件下或者变性条件下被洗脱，从而被分离纯化。 本产品对His标签重组蛋白的亲和力强、选择性高、结合容量大，使用本产品可以获得高纯度的His标签重组蛋白，可用于简单、快速、高效地纯化细菌、哺乳动物细胞、昆虫及杆状病毒等多种表达系统中表达的His标签重组蛋白。纯化的蛋白可以用于结构和功能研究、抗体制备、蛋白与蛋白相互作用、蛋白与核酸相互作用等方面的研究。 本产品采用了一种高度交联的6%琼脂糖凝胶为基质，并使用了进一步优化的接臂和镍离子螯合技术，和目前市售的同类产品相比，非特异性蛋白结合显著降低。 本产品可以耐受8M尿素或6M盐酸胍，可用于非变形条件和变性条件下His标签蛋白的纯化。DTT、β-巯基乙醇等还原剂会还原螯合结合的镍离子，而EDTA/EGTA等二价离子强螯合剂则会使本产品脱镍，因此在蛋白样品或者纯化过程所使用的溶液中不能含有还原剂和螯合剂。如需在强还原剂如DTT/β-巯基乙醇或螯合剂EDTA/EGTA存在的条件下纯化蛋白，推荐使用阿拉丁的耐受还原剂和螯合剂。 本产品已经螯合了镍离子，呈蓝绿色，凝胶的颗粒直径为45-165μm。可耐受的最大压力为0.3MPa，约合43.5psi。采用固定流速进行蛋白纯化时的最大流速为300cm/h。 本产品可以高容量、高特异性地结合His标签蛋白。最大蛋白结合量约为25-35mg蛋白(分子量为55kD的蛋白)/毫升凝胶。实际使用时的最大结合量主要取决于待纯化的His标签重组蛋白的分子量大小，分子量越大则最大结合容量越大，分子量越小则最大结合容量越小。但对于分子量相同的蛋白，也会因为蛋白本身特性的不同，结合的最大容量也会有所不同。 本产品储存在20%乙醇中。本产品包装体积中50%为凝胶，例如10ml总体积中5ml为凝胶，5ml为液体。使用时宜把凝胶充分重悬后再吸取。 每10ml包装的本产品最多可纯化100-200mg带有His标签的重组蛋白(分子量为55kD的蛋白)，分子量更大的蛋白结合容量会更大一些。
注意事项： 请勿在-20℃或更低温度冷冻保存本产品。 本产品使用过程中，缓冲试剂如Tris、HEPES、MOPS等的浓度不宜超过100mM，SDS和sarkosyl的浓度不宜超过0.3%，Triton X-100、Tween-20、NP-40的浓度不宜超过2%，脱氧胆酸钠、CHAPS的浓度不宜超过1%，组氨酸浓度不宜超过20mM，钙离子浓度不宜超过5mM，钠离子和镁离子浓度可以高达2M，盐酸胍浓度可以高达6M，尿素浓度可以高达8M，甘油浓度可以高达50%。本产品不能耐受还原剂、螯合剂和强酸强碱。其它未提及试剂的兼容性可以参考上述试剂，但还有待实验验证。 保存和纯化过程中应始终保持凝胶湿润。 若离心不能完全除去蛋白样品中的不溶物，可以将样品溶液用0.45μm的滤膜过滤。 蛋白样品收集后宜尽快完成纯化工作，并应始终放置在4℃或冰浴，以减缓蛋白降解。为有效抑制蛋白降解，可以在裂解液中添加适量的蛋白酶抑制剂混合物。 若使用本说明书提供的条件无法达到理想的纯化效果，可尝试改变洗涤液和洗脱液中咪唑的浓度和(或)pH，以达到最优效果。 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：
常见问题：
相关溶液配制方法：
Reductant and Chelator-resistant: Aladdin's UltraBio™ His-tag Purification Resin (Reductant&Chelator-resistant) provides a simple, rapid, and highly efficient method for purifying His-tagged proteins from cell lysates, with high specificity. It is compatible with a certain concentration of reductants and chelants. A His-tag is a string of histidine residues, typically fused to the N or C terminus of a recombinant protein. The classic his-tag comprises 6 histidine residues, named hexahistidine tag (6xHis-tag). Some recombinant proteins are engineered to have two hexahistidine tags. The His-tag has a high affinity for nickel ions (Ni2+) which is immobilized on UltraBio™ His-tag Purification Resin. Therefore, the His-tag Purification Resin can be used to purify His-tagged recombinant proteins from a crude lysate, while most other proteins in the lysate will not bind to the resin, or bind only weakly. UltraBio™ His-tag Purification Resin has a strong affinity for His-tagged proteins, with high specificity and binding capacity, which enables simple, rapid, and efficient purification of the expressed His-tagged protein from a broad host such as bacteria, mammals, insects, Baculovirus, or yeast. The obtained His-tagged protein with high purity can be used for protein structure and function analysis, antibody preparation, protein interaction studies, protein-nucleic acid interaction studies, etc. UltraBio™ His-tag Purification Resin is based on highly crossed-linked 6% agarose gel on which nickel ions are immobilized by the optimized arm coupling and chelating technology. Compared to other products with similar functions, UltraBio™ His-tag Purification Resin exhibits lower non-specificity, stronger resistance to pressure, and tighter binding Ni2+ to the resin. This product has minimal leaching of Ni2+ and can be reused many times without reloading nickel ions. Ni-NTA agarose and most other nickel resin products are not compatible with commonly used concentrations of reducing agents, such as dithiothreitol (DTT) and β -mercaptoethanol, and chelating agents, such as EDTA. However, this product can be compatible with 20mM DTT and 20mM EDTA, facilitating the purification of His-tagged proteins from samples containing reductants and/or chelants. This product is not compatible with 8M urea and 6M guanidine hydrochloride. To purify proteins from inclusion bodies that require 8M urea or 6M guanidine hydrochloride for protein solubilization, dialysis should be performed to remove urea and guanidine hydrochloride before using this product for protein purification. UltraBio™ His-tag Purification Resin is a ready-to-use Ni2+ chelate blue resin, with a particle diameter of 45-165μm. It can tolerate pressure up to 0.025MPa (~5.8psi). The recommended flow rate is 0.5ml/min when using a fixed flow rate for protein purification. UltraBio™ His-tag Purification Resin binds His-tagged protein with high capacity and high specificity. Per milliliter gel can bind up to 20-30mg of His-tagged protein. The maximum binding capacity in actual use depends on the molecular weight and characteristics of the protein to be purified. The larger the molecular weight, the greater the maximum binding capacity, and vice versa. For proteins with the same molecular weight, the maximum binding capacity of this resin might be different due to different characteristics of the proteins. This product is stored in 30% ethanol. A total volume of 10ml contains 5ml of gel and 5ml of liquid. It is advisable to fully resuspend the gel prior to use. Per 10ml of this product can purify up to 100-150mg of His-tagged recombinant protein. Precautions： Store the resin at 4ºC upon receipt. Do NOT freeze. During the use of this product, the concentration of buffer reagents such as Tris, HEPES, and MOPS, should not exceed 100mM, EDTA should not exceed 20mM, SDS and sarkosyl should not exceed 0.3%, and the concentration of Triton, Tween, and NP-40 should not exceed 2%. The concentration of sodium deoxycholate and CHAPS should not exceed 1%, histidine should not exceed 20mM, calcium ions should not exceed 5mM, sodium and magnesium ions can be up to 2M, and glycerol can be up to 50%. The compatibility of other reagents not mentioned is yet to be tested. Keep the gel moist during storage or purification. A 0.45μm filter membrane can be used to remove particulates from cell lysates that could not be completely removed by centrifuge. Protein samples should be purified after collection as soon as possible and kept at 4ºC or on ice to minimize protein degradation or denaturation. Protein degradation can also be inhibited by the addition of Protease Inhibitor Cocktail. If protein purification cannot be achieved ideally when using the conditions provided in this manual, optimize imidazole concentration and/or the pH of wash and elution buffers. 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. Fast Flow;Resistant to Reductants and Chelators: Aladdin's UltraBio™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant) provides a simple, rapid, highly efficient and specific method for purifying His-tagged proteins from cell lysates, with fast flow rate. This new developed resin is high-pressure tolerant and is compatible with reducing and chelating agents. A His-tag is a string of histidine residues, typically fused to the N or C terminus of a recombinant protein. The classic his-tag comprises 6 histidine residues, named hexahistidine tag (6xHis-tag). Some recombinant proteins are engineered to have two hexahistidine tags. The His-tag has a high affinity for nickel ions (Ni2+) which is immobilized on UltraBio™ His-tag Purification Resin. Therefore, the UltraBio™ His-tag Purification Resin can be used to purify His-tagged recombinant proteins from a crude lysate, while most other proteins in the lysate will not bind to the resin, or bind only weakly. This product has high affinity for His-tagged proteins, with high specificity and binding capacity, which enables simple, rapid, and efficient purification of the expressed His-tagged protein from a broad host such as bacteria, mammals, insects, Baculovirus, or yeast. The obtained His-tagged protein with high purity can be used for protein structure and function analysis, antibody preparation, protein interaction studies, protein-nucleic acid interaction studies, etc. his product employs the highly cross-linked 6% agarose gel on which nickel ions are immobilized by the optimized arm coupling and chelating technology. Compared to other products with similar functions, UltraBio™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant) exhibits lower non-specificity, stronger resistance to pressure, and tighter binding Ni2+ to the resin. This product almost has no leaching of Ni2+ and can be reused many times without the need of reloading nickel ions. Ni-NTA agarose and most similar nickel column products cannot tolerate common concentrations of reducing agents such as dithiothreitol (DTT), β-mercaptoethanol, etc., as well as common concentrations of chelating agents such as EDTA, etc., in protein samples; however, this product can well tolerate reducing agents such as DTT and chelating agents such as EDTA. This product can tolerate 0.01 M HCl, 0.01 M NaOH (test condition: 40°C, 7d); 10 mM EDTA, 5 mM DTT, 5 mM TCEP, 20 mM β-Mercaptoethanol, 1 M NaOH, 4 M Guanidine hydrochloride (test condition: 25°C, 24h) ; 500 mM imidazole, 100 mM EDTA (test conditions: 25℃, 2 h); 30% Isopropanol (test conditions: 25℃, 20 min). This offers the possibility of purifying protein samples that require the addition of reducing or chelating agents. This product is a ready-to-use Ni2+ chelate blue resin, with a particle diameter of 45-165μm. It can tolerate pressure up to 0.3MPa (~43.5 psi). The recommended flow rate is 600cm/h when using a fixed flow rate for protein purification. UltraBio™ His-tag Purification Resin binds His-tagged protein with high capacity and high specificity. Per milliliter gel can bind up to 15 mg of His-tagged protein. The maximum binding capacity in actual use depends on the molecular weight and characteristics of the protein to be purified. The larger the molecular weight, the greater the maximum binding capacity, and vice versa. For proteins with the same molecular weight, the maximum binding capacity of this resin might be different due to different characteristics of the proteins. This product is stored in 20% ethanol. A total volume of 10ml contains 5ml of gel and 5ml of liquid. Precautions: Store at 2-8 ℃. Do NOT freeze. This product is for RUO only. Not for drug, household, or other uses. For your safety and health, please wear a lab coat and disposable gloves during the operation. Denaturant- resistant: Aladdin's UltraBio™ His-tag Purification Resin (Denaturant-resistant) provides a simple, rapid, and highly efficient method for purifying His-tagged proteins from cell lysates under either native or denaturing conditions, with high specificity. This product is compatible with strong denaturants, such as urea and guanidine hydrochloride. A His-tag is a string of histidine residues, typically fused to the N or C terminus of a recombinant protein. The classic His-tag comprises 6 histidine residues, named hexahistidine tag (6xHis-tag). Some recombinant proteins are engineered to have two hexahistidine tags. His-tag has a high affinity for nickel ions (Ni2+) which is immobilized on UltraBio™ His-tag Purification Resin. Therefore, the UltraBio™ His-tag Purification Resin can be used to purify His-tagged recombinant proteins from a crude lysate, while most other proteins in the lysate will not bind to the resin, or bind only weakly. UltraBio™ His-tag Purification Resin has a strong affinity for His-tagged proteins, with high specificity and binding capacity, which enables simple, rapid, and efficient purification of His-tagged protein from a broad host such as bacteria, mammals, insects, Baculovirus, or yeast. The obtained His-tagged protein with high purity can be used for protein structure and function analysis, antibody preparation, protein interaction studies, protein-nucleic acid interaction studies, etc. UltraBio™ His-tag Purification Resin is based on highly crossed-linked 6% agarose gel on which nickel ions are immobilized by the optimized arm coupling and chelating technology. Compared to other products with similar functions, UltraBio™ His-tag Purification Resin exhibits much lower non-specificity. UltraBio™ His-tag Purification Resin (Denaturant-resistant) is compatible with 8M urea and 6M guanidine hydrochloride, but not reductants and chelators. Reductants, such as dithiothreitol (DTT) and β -mercaptoethanol, and chelating agents, such as EDTA and EGTA, will cause the leakage of Ni2+. UltraBio™ His-tag Purification Resin is a ready-to-use Ni2+ chelate blue resin, with a particle diameter of 45-165μm. It can tolerate pressure up to 0.025MPa (~5.8psi). The recommended flow rate is 0.5ml/min when using a fixed flow rate for protein purification. UltraBio™ His-tag Purification Resin binds His-tagged protein with high capacity and high specificity. Per milliliter of gel can bind up to 25-35mg of His-tagged protein. The maximum binding capacity in actual use depends on the molecular weight and characteristics of the protein to be purified. The larger the molecular weight, the greater the maximum binding capacity, and vice versa. For proteins with the same molecular weight, the maximum binding capacity of this resin might be different due to different characteristics of the proteins. This product is stored in 20% ethanol. A total volume of 10ml contains 5ml of gel and 5ml of liquid. It is advisable to fully resuspend the gel prior to use. 10ml of this product can purify up to 100-200mg of His-tagged recombinant protein with a molecular weight of 55kD. Precautions： Store the resin at 4℃ upon receipt. Do NOT freeze. During the use of this product, the concentration of buffer reagents such as Tris, HEPES, and MOPS, should not exceed 100mM. The concentrations of Triton X-100, Tween-20, and NP-40 should not exceed 2%. The concentration of sodium deoxycholate and CHAPS should not exceed 1%, histidine should not exceed 20mM, calcium ions should not exceed 5mM, sodium and magnesium ions can be up to 2M, and glycerol can be up to 50%. This product is compatible with urea up to 8M, and guanidine hydrochloride up to 6M, but not compatible with reductants, chelating agents, or strongly acidic or alkaline solutions. The compatibility of other reagents not mentioned is yet to be tested. Keep the gel moist during storage or purification. A 0.45μm filter membrane can be used to remove particulates from cell lysates that could not be completely removed by centrifuge. Protein samples should be purified after collection as soon as possible and kept at 4℃ or on ice to minimize protein degradation or denaturation. Protein degradation can also be inhibited by the addition of Protease Inhibitor Cocktail. If protein purification cannot be achieved ideally when using the conditions provided in this manual, optimize imidazole concentration and/or the pH of wash and elution buffers. 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. Fast Flow;Denaturant-resistant: Aladdin's UltraBio™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) provides a simple, rapid, highly efficient and specific method for purifying His-tagged proteins from cell lysates, with fast flow rate. This new developed resin is high-pressure tolerant and is compatible with strong protein denaturants such as urea and guanidine hydrochloride. A His-tag is a string of histidine residues, typically fused to the N or C terminus of a recombinant protein. The classic his-tag comprises 6 histidine residues, named hexahistidine tag (6xHis-tag). Some recombinant proteins are engineered to have two hexahistidine tags. The His-tag has a high affinity for nickel ions (Ni2+) which is immobilized on UltraBio™ His-tag Purification Resin. Therefore, the UltraBio™ His-tag Purification Resin can be used to purify His-tagged recombinant proteins from a crude lysate, while most other proteins in the lysate will not bind to the resin, or bind only weakly. Store at 4℃ for up to 1 year. This product has high affinity for His-tagged proteins, with high specificity and binding capacity, which enables simple, rapid, and efficient purification of the expressed His-tagged protein from a broad host such as bacteria, mammals, insects, Baculovirus, or yeast. The obtained His-tagged protein with high purity can be used for protein structure and function analysis, antibody preparation, protein interaction studies, protein-nucleic acid interaction studies, etc. This product employs the highly cross-linked 6% agarose gel on which nickel ions are immobilized by the optimized arm coupling and chelating technology. Compared to other products with similar functions, UltraBio™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant) exhibits lower non-specificity, stronger resistance to pressure, and tighter binding Ni2+ to the resin. This product can tolerate 8M urea or 6M guanidine hydrochloride and can be used for the purification of His-tagged proteins under native or denaturing conditions. Reducing agents such as DTT and β-mercaptoethanol will reduce chelated nickel ions, while strong chelating agents such as EDTA/EGTA and other divalent ions will de-nickel the product. Therefore, reducing and chelating agents should not be contained in the solution used for protein purification. For protein purification in the presence of strong reducing agents such as DTT/β-mercaptoethanol or chelating agents such as EDTA/EGTA. This product is a ready-to-use Ni2+ chelate blue resin, with a particle diameter of 45-165μm. It can tolerate pressure up to 0.3MPa (~43.5 psi). The recommended flow rate is 0.5ml/min when using a fixed flow rate for protein purification. This product binds His-tagged proteins with high capacity and specificity. The maximum protein binding capacity is 25-35mg protein (with a molecular weight of 55kD)/ml gel. The maximum binding capacity in practice depends on the molecular weight and characteristics of the His-tagged recombinant protein to be purified. The larger the molecular weight, the higher the maximum binding capacity, and vice versa. However, for proteins with the same molecular weight, the maximum binding capacity may vary depending on the properties of the purified protein. This product is stored in 20% ethanol. A total volume of 10ml of this product contains 5ml of gel and 5ml of liquid. It is advisable to fully resuspend the gel prior to use. Up to 100-200mg of His-tagged recombinant protein (with a molecular weight of 55kD) can be purified per 10ml of this product. Precautions： Store the resin at 4℃ upon receipt. Do NOT freeze. During the use of this product, the concentration of buffer reagents such as Tris, HEPES, and MOPS, should not exceed 100mM. The concentrations of Triton X-100, Tween-20, and NP-40 should not exceed 2%. The concentration of sodium deoxycholate and CHAPS should not exceed 1%, histidine should not exceed 20mM, calcium ions should not exceed 5mM, sodium and magnesium ions can be up to 2M, and glycerol can be up to 50%. This product is compatible with urea up to 8M and guanidine hydrochloride up to 6M, but not compatible with reductants, chelating agents, or strongly acidic or alkaline solutions. The compatibility with other reagents not mentioned is yet to be tested. Keep the gel moist during storage or purification. A 0.45μm filter membrane can be used to remove particulates from cell lysates that could not be completely removed by centrifuge. Protein samples should be purified after collection as soon as possible and kept at 4℃ or on ice to minimize protein degradation or denaturation. Protein degradation can also be inhibited by the addition of Protease Inhibitor Cocktail. If protein purification cannot be achieved ideally when using the conditions provided in this manual, optimize imidazole concentration and/or the pH of wash and elution buffers. 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. |
储存与运输
| 储存温度 | 2-8°C储存 |
|---|---|
| 运输条件 | 冰袋运输 |
| 稳定性与储存 | 4℃保存，至少一年有效。 |
质检证书（CoA,COO,BSE/TSE 和分析图谱)
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ24F1114387 |
分析证书 | H752721 |