BioReagent,for IP 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
2-8°C储存,禁止冷冻。低温运输,禁止冷冻 。请查阅批次 COA 获取详细规格。
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
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。
阿拉丁生产的UltraBio™ Anti-MBP Agarose(UltraBio™ Anti-MBP琼脂糖凝胶)，由高品质的MBP抗体与4%琼脂糖凝胶共价偶联制成，本产品可特异性地与动植物或者微生物裂解液、血清、腹水等样品中含有MBP标签的蛋白结合，从而用于带有MBP标签的融合蛋白或蛋白复合物的免疫沉淀(Immunoprecipitation, IP)或免疫共沉淀(Co-IP)。麦芽糖结合蛋白(Maltose binding protein, MBP)，是大肠杆菌(E. coli)麦芽糖(Maltose)/麦芽糊精(Maltodextrin)系统的一部分，由malE基因编码，负责麦芽糊精的摄取、分解和代谢，分子量约为42kDa。MBP标签，即麦芽糖结合蛋白标签，可融合在蛋白的N端或C端，通常融合在N端。MBP标签具有以下优点：能增加外源蛋白的可溶性；适用范围广；可促进蛋白的正确折叠；高特异性，纯化方便且温和；MBP标签作为标签蛋白，后续通过MBP抗体、Anti-MBP琼脂糖凝胶即可对目的蛋白的表达、定位及功能进行检测或对目的蛋白进行纯化、免疫沉淀或免疫共沉淀等。
阿拉丁UltraBio™ Anti-MBP琼脂糖凝胶储存在含0.02% ProClin 300的PBS中，凝胶和保护液的体积比为1：1（即50%凝胶悬液，每毫升本产品中共含有0.5ml凝胶沉淀物），我司产品规格为总体积。产品信息见下表：
|
使用说明
1、样品准备
1.1上柱之前要确保样品溶液有合适的离子强度和pH值，可以用平衡液对样品或细胞培养液稀释，或者用平衡液透析。
1.2样品在上样前建议离心或用0.22μm或0.45μm滤膜过滤，减少杂质，提高蛋白纯化效率和防止堵塞柱子。
1.3选择合适的裂解液，用于制备细胞或组织的裂解液。优先推荐选择阿拉丁生产的P0013 Western及IP细胞裂解液用于细胞或组织样品的裂解。根据特定的实验目的，如有必要，也可以使用阿拉丁生产的P0013B RIPA裂解液(强)、P0013C RIPA裂解液(中)或P0013D RIPA裂解液(弱)用于样品的制备。如果使用自行配制的或其它公司生产的裂解液，需要确保裂解液的pH为6-8。
2、缓冲液的准备
所用水和缓冲液在使用之前建议用0.22μm或0.45μm滤膜过滤。
平衡液：0.01M PBS，pH7.4
洗杂液：0.01M PBS，0.05% Tween-20，pH7.4
3、免疫沉淀操作流程
(1) 填料准备：取40µL的Anti-MBP琼脂糖凝胶(凝胶实际体积20 µL) 混合液加入到1.5mL 离心管中，5000×g离心1min，吸弃上清；
(2) 填料加入0.5mL平衡液，悬浮填料（使填料处于与目的蛋白相同的缓冲体系下，起到保护蛋白的作用），5000×g离心1min，吸弃上清。重复一次。
(3) 加入300µL样品裂解液到处理好的填料中，混合均匀，在室温下置于翻转混合仪轻轻翻转离心管，促使样品和填料充分接触并吸附，室温反应3h。5000×g离心1min，吸弃上清。
(4) 洗杂：加入0.5mL的洗杂液，悬浮填料，轻轻混匀，5000×g离心1min，吸弃上清。再重复三次。确保去除非特异性吸附。
(5)变性洗脱：加入100µL 1×Loading Buffer，95℃加热5min。5000×g离心1min，吸取上清进行SDS-PAGE电泳检测。
Note：实验室常规蛋白上样缓冲液（Loading Buffer）中含有β-巯基乙醇和DTT，可以使填料中抗体重链和轻链断开。含有SDS的样品缓冲液可以使Anti-MBP抗体变性，洗脱后的Anti-MBP琼脂糖凝胶不能重复使用。
注意事项
1. 本产品使用前一定要充分重悬，即充分颠倒若干次使混合均匀。
2. 在免疫沉淀时，建议设置阳性和阴性对照组。
3. 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，以减缓蛋白降解或变性。
4. 高浓度的DTT、巯基乙醇、盐酸胍等对本产品的非特异性结合可能有一定影响，但Western及IP细胞裂解液、RIPA裂解液或NP-40裂解液等都完全适用。
5. 为有效抑制蛋白降解，可以在蛋白样品中添加适量的蛋白酶抑制剂混合物，例如阿拉丁的P1005/P1006蛋白酶抑制剂混合物(通用型)、P1048/P1049蛋白酶磷酸酶抑制剂混合物(通用型, 质谱兼容, 50X)、P1010/P1011蛋白酶抑制剂混合物(哺乳动物样品抽提用, 100X)、P1050/P1051蛋白酶磷酸酶抑制剂混合物(哺乳动物样品抽提用, 50X)等，或P1025/P1026蛋白酶抑制剂混合物(细菌抽提用)。
6. 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。
7. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
Aladdin's UltraBio™ Anti-MBP Agarose is produced by high-quality MBP antibodies to 4% agarose gel. These agarose can specifically bind MBP fusion proteins in lysates, serum, ascites, or other samples from animals, plants, or microorganisms. This product is mainly used for immunoprecipitation (IP), co-immunoprecipitation (Co-IP), or purification of MBP-tagged proteins or their protein complexes. Maltose binding protein (MBP) encoded by the malE gene is a component of the E. coli maltose/maltodextrin system and regulates the uptake and catabolism of maltodextrins, with a molecular weight of approximately 42kDa [1]. The MBP tag can be fused to the N- or C-terminus of a protein, with the N-terminus being the most common fusion site. The MBP tag has several advantages: 1. Increased solubility of heterologous proteins.2. Wide applicability in different hosts/systems.3. Facilitation of proper protein folding. 4. High specificity allowing for mild and convenient purification. 5. Versatile tool for downstream analysis: Following fusion, the expression, localization, and function of the target protein can be analyzed, or the protein can be purified, immunoprecipitated (IP), or co-immunoprecipitated (Co-IP) using MBP-specific antibodies or Anti-MBP Agarose gel.
Aladdin UltraBio™ Anti-MBP Agarose is stored in PBS containing 0.02% ProClin 300, with a settled gel to storage solution ratio of 1:1(This product is a 50% gel suspension, containing 0.5 ml of settled agarose per milliliter. Our product specifications are based on total volume. Product information is detailed in the table below:
|
Instruction for use
1. Sample Preparation
1.1 Ensure the sample solution has appropriate ionic strength and pH before loading. Dilute the sample or cell culture supernatant with equilibration buffer, or dialyze the sample against equilibration buffer.
1.2 Clarify the sample by centrifugation or filtration through a 0.22 μm or 0.45 μm membrane to reduce impurities, improve purification efficiency, and prevent column clogging.
1.3 Preparation of protein samplesa.Lysis cells or tissues with appropriate lysis buffer, such as ’s Cell Lysis Buffer for Western and IP . Under certain circumstances, ’s RIPA Strong Lysis Buffer , RIPA Medium Lysis Buffer , or RIPA Weak Lysis Buffer can be used. Other lysis buffers with a pH6-8 can also be used
2. Buffer Preparation
It is recommended to filter water and buffers through a 0.22 μm or 0.45 μm membrane before use.
Equilibration Buffer: 0.01M PBS，pH7.4；
Wash Buffer: 0.01M PBS，0.05% Tween-20，pH7.4
3. Immunoprecipitation (IP) Procedure
3.1 Resin Preparation: Add 40 µL of Anti-MBP Agarose suspension (20 µL settled agarose) to a 1.5 mL tube. Centrifuge at 5,000 × g for 1 min. Discard the supernatant.
3.2 Add 0.5 mL of Equilibration Buffer to resuspend the agarose (Place the agarose in the same buffer system as the target protein to protect the protein.). Centrifuge at 5,000 × g for 1 min. Discard the supernatant. Repeat this wash step once.
3.3 Add 300 µL of sample lysate to the prepared agarose from step 3.2. Mix and incubate on a tube rotator at room temperature for 3 hours. Centrifuge at 5,000 × g for 1 min. Discard the supernatant.
3.4 Washing: Add 0.5 mL of Wash Buffer, resuspend the agarose, and mix gently. Centrifuge at 5,000 × g for 1 min. Discard the supernatant. Repeat this wash step three more times.
3.5 Denaturing Elution (SDS-PAGE): Add 100 µL of 1× Loading Buffer to the agarose. Heat at 95°C for 5 min. Centrifuge at 5,000 × g for 1 min, and load the supernatant directly onto an SDS-PAGE gel for analysis.
Note: The Standard loading buffer used in laboratories contains β-mercaptoethanol and DTT, which can break the disulfide bonds between the antibody heavy and light chains on the agarose. This method denatures the antibody, rendering the resin unusable for reuse.
Matters needing attention
1. This product must be fully resuspended by inverting the tube several times prior to use.
2. For immunoprecipitation assays, it is recommended to include both positive and negative controls.
3. Protein samples should be purified as soon as possible after collection and should always be placed at 4℃ or on ice to minimize protein degradation or denaturation.
4. The non-specific binding capability of this product can be affected by high concentrations of DTT, mercaptoethanol, and guanidine hydrochloride, but it is applicable in Cell lysis buffer for Western and IP , RIPA Lysis Buffer , and NP-40 Lysis Buffer .
5. Protein degradation can also be inhibited by adding appropriate protease inhibitors, such as Protease Inhibitor Cocktail for General Use , Protease and Phosphatase Inhibitor Cocktail for General Use (MS-safe, 50X) , Protease Inhibitor Cocktail for Mammalian Cell and Tissue Extracts , Protease and Phosphatase Inhibitor Cocktail for Mammalian Cell and Tissue Extracts , etc.
6. This product is for R&D only. Not for drug, household, or other uses.
7. For your safety and health, please wear a lab coat and disposable gloves during the operation.
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0434639 |
分析证书 | U1508192 |
| ZJ26F0434638 |
分析证书 | U1508192 |
危险品化学品经营许可证（带存储）