- ￥1290 - 3960
- 帛科
- BK-DB6327
- 国产
- 2025年07月15日
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- 详细信息
- 文献和实验
- 技术资料
- 库存：
31
- 英文名：
Recombinant Human mIL15 protein
- 保质期：
12个月
- 供应商：
上海帛科
- 保存条件：
-20 to -80 °C
- 规格：
50μg、100ug 、1mg
| 货号 | BK-DB6327 | 用途 | 仅供科研研究实验 |
| 纯度 | >90% as determined by SDS-PAGE | 预测分子量 | 14.62 kDa |
| 表达宿主 | E.coli | 种属 | Homo sapiens (Human) |
蛋白构建: A DNA sequence encoding the human mIL15 (Met21-Gly133) was fused with His tag
制剂: Supplied as solution form in PBS pH 7.5 or lyophilized from PBS pH 7.5.
运输方式: In general, proteins are provided as lyophilized powder/frozen liquid. They are shipped out with dry ice/blue ice unless customers require otherwise.
稳定性&储存: Use a manual defrost freezer and avoid repeated freeze thaw cycles.Store at 2 to 8 °C for one week .Store at -20 to -80 °C for twelve months from the date of receipt.
复溶: Reconstitute in sterile water for a stock solution.A copy of datasheet will be provided with the products, please refer to it for details.
分子别名: Interleukin-5,IL-5,B-cell growth factor II,BCGF-II,Cytotoxic T-lymphocyte inducer,Eosinophil differentiation factor,T-cell replacing factor,TRF.
背景介绍: Interleukin 5 (IL-5) is a member of the interleukin family with length of 115 amino acids. Interleukins are a group of cytokines (secreted proteins / signaling molecules) that were first seen to be expressed by white blood cells (leukocytes) and has been found in a wide variety of body cells. Interleukin 5 or IL-5 is produced by T helper-2 cells and mast cells. It helps to stimulate B cell growth and increase immunoglobulin secretion and is considered as a key mediator in eosinophil activation. Interleukin 5 (IL-5) has long been associated with several allergic diseases, including allergic rhinitis and asthma. Growth in the number of circulating, airway tissue, and induced sputum eosinophils have been oserved in patients with these diseases. IL-5 also had something with the terminally differentiated granulocyte eosinophils. IL-5 was originally found as an eosinophil colony stimulating factor. It has been proved to be a major regulator of eosinophil accumulation in tissues, and can modulate eosinophil behavior at every stage from maturation to survival.
重组蛋白质的可溶性鉴定：
1. 将按上法诱导培养后收集的菌体重悬于裂解液 1 (Lysis buffer under native conditions )中，然后在－80 oC 低温冰箱中放置 10 min。
2. 冰中解冻。
3. 在冰浴上用超声破碎仪破菌 6 次， 每次 10 sec，间歇 10 sec，电压 200-300 V。
4. 10000g，4oC，离心 20min，取上清（为溶液 A）， － 20 oC 保存； 另将沉淀用 同样裂解液 1 溶解（为溶液 B），同样 －20 oC 保存，供后继分析使用。
5. 将上述 A、B 溶液和诱导前后的细菌进行 SDS-PAGE 电泳， 考马斯亮蓝染色， 比较分析重组蛋白质的溶解性。如果诱导表达的蛋白质位于 A 溶液中， 则为可溶性蛋白；如果是在 B 溶液中，则为非可溶蛋白。
重组蛋白质的分离纯化：
1. 将菌体沉淀溶于适量裂解液 2（Lysis buffer under denaturing conditions ）中，室温下搅拌和吹打沉淀，避免泡沫生成。
2. 10000g ，4 oC，离心 30 min，收集上清液。
3. 将 Ni-NTAAgarose 充填柱子，并连接于 Pharmarcia 低压液相层析系统，用 5 倍柱体积的裂解液 2 平衡 Ni-NTAAgarose，调节 A280 值至零线。
4. 将适量上清液上样到 Ni-NTAAgarose 柱子中， 并用 lysis buffer 冲洗至 A280 值低于 0.01。
5. 分别用 5 ～ 10 倍柱体积的清洗液 1 和清洗液 2（Washbuffer 1 and 2）清洗柱 子，直至 A280 值低于 0.01。
6. 用洗脱液（Elution buffer）洗脱重组蛋白质， 在 A280 值监测下，收集出现峰 线后含有重组蛋白的所有洗脱液。
表达载体在基因工程有以下几种元件：
（1）选择标志的编码序列；
（2）可控转录的启动子；
（3）转录调控序列(转录终止子，核糖体结合位点)；
（4）一个多限制酶切位点接头；
（5）宿主体内自主复制的序列。
重组蛋白质的诱导表达：
1. 挑取转化有质粒的单菌落， 接种于 3ml 选择性 LB 液体培养基中， 37 oC，250 rpm/min 振摇培养过夜。
2. 次日将培养过夜的菌液 500 μl 再接种于 10ml（1:20）选择性 LB 液体培养基 中， 37 oC，250 rpm/min 振摇培养至光密度（OD600=0.6）时，取 1 ml 样本作 为诱导前标本， 10000g 离心 1min 收集菌体沉淀，－20 oC 冻存备用。
3. 加入 1 mol/L IPTG 于菌液中，使 IPTG 终浓度为 1 mM ，37 oC ，250 rpm/min 振摇培养 4 ～ 5 小时。取 1 ml 样本作为诱导后标本，同上法收集菌体沉淀， -20 oC 冻存备用。
4. 将诱导前后菌体沉淀用 20 ~ 40 μl PBS（pH= 8.0）重悬， 加入等体积的 2×SDS 上样缓冲液， 煮沸加热 5min，SDS 聚凝胶（SDS-PAGE）电泳分离 ， 考马斯亮蓝染色 3 小时后，脱色观察结果。
5. 选取诱导成功的细菌克隆，扩大诱导规模，收集菌体沉淀，于－ 20 oC 保存，准备做下一步分析及纯化。
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| MCR8蛋白封闭多肽 | NR5A2 Antibody Blocking Peptide |
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| 破伤风毒素重链封闭多肽 | Recombinant Human mIL15 proteinDRD2 Antibody Blocking Peptide |
| KIF1B Antibody Blocking Peptide | |
| 软骨基质蛋白封闭多肽 | OGFRL1 Antibody Blocking Peptide |
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