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- 帛科
- BK-DB3049
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- 详细信息
- 文献和实验
- 技术资料
- 库存：
46
- 英文名：
Recombinant Human MMP12 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
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。
商品介绍：
内毒素: Please contact with the lab for this information.
蛋白构建: A DNA sequence encoding the human MMP12 (Ala290-Gly469) 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: Macrophage metalloelastase,MME,Macrophage elastase,ME,hME,Matrix metalloproteinase-12,MMP-12.
背景介绍: Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade components of the extracellular matrix (ECM) and play essential roles in various physiological processes such as morphogenesis, differentiation, angiogenesis and tissue remodeling, as well as pathological processes including inflammation, arthritis, cardiovascular diseases, pulmonary diseases and tumor invasion. Macrophage metalloelastase, also known as Matrix metalloproteinase-12, Macrophage elastase, MMP12, and MMP-12, is a secreted protein which belongs to thepeptidase M1A family. MMP12 is a macrophage-secreted elastase that is highly induced in the liver and lung in response to S. mansoni eggs and contains four hemopexin-like domains. MMP12 is a proteolytic enzyme responsible for cleavage of plasminogen to angiotensin, which has an angiostatic effect. It may be involved in tissue injury and remodeling and has significant elastolytic activity. It may be related to prognosis in breast cancer patients. MMP12 promotes fibrosis by limiting the expression of specific ECM-degrading MMPs. Like MMP12, MMP13 expression is highly dependent on IL-13 and type II-IL-4 receptor signaling. MMP12 is a potent proinflammatory and oncogenic molecule. MMP12 up-regulation plays a critical role in emphysema to lung cancer transition that is facilitated by inflammation.
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。
公司出售的相关产品：
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。
商品介绍：
| 货号 | BK-DB3049 | 用途 | 仅供科研研究实验 |
| 纯度 | >90% as determined by SDS-PAGE | 预测分子量 | 22.5 kDa |
| 表达宿主 | E.coli | 种属 | Homo sapiens (Human) |
蛋白构建: A DNA sequence encoding the human MMP12 (Ala290-Gly469) 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: Macrophage metalloelastase,MME,Macrophage elastase,ME,hME,Matrix metalloproteinase-12,MMP-12.
背景介绍: Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade components of the extracellular matrix (ECM) and play essential roles in various physiological processes such as morphogenesis, differentiation, angiogenesis and tissue remodeling, as well as pathological processes including inflammation, arthritis, cardiovascular diseases, pulmonary diseases and tumor invasion. Macrophage metalloelastase, also known as Matrix metalloproteinase-12, Macrophage elastase, MMP12, and MMP-12, is a secreted protein which belongs to thepeptidase M1A family. MMP12 is a macrophage-secreted elastase that is highly induced in the liver and lung in response to S. mansoni eggs and contains four hemopexin-like domains. MMP12 is a proteolytic enzyme responsible for cleavage of plasminogen to angiotensin, which has an angiostatic effect. It may be involved in tissue injury and remodeling and has significant elastolytic activity. It may be related to prognosis in breast cancer patients. MMP12 promotes fibrosis by limiting the expression of specific ECM-degrading MMPs. Like MMP12, MMP13 expression is highly dependent on IL-13 and type II-IL-4 receptor signaling. MMP12 is a potent proinflammatory and oncogenic molecule. MMP12 up-regulation plays a critical role in emphysema to lung cancer transition that is facilitated by inflammation.
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。
公司出售的相关产品：
| Recombnant Human GRK6 proten | VAMP-3 Antibody Blocking Peptide |
| Recombinant Human REXO2 | GPR182 Antibody Blocking Peptide |
| CDK11 Antibody Blocking Peptide | TNFSF14 Antibody Blocking Peptide |
| BEND2 Antibody Blocking Peptide | FAM78B Antibody Blocking Peptide |
| phospho-B MyB (Thr487) Antibody Blocking Peptide | CLDN25 Antibody Blocking Peptide |
| GKRP Antibody Blocking Peptide | Phospho-ERBB3 (Tyr1289) Antibody Blocking Peptide |
| MRPL24 Antibody Blocking Peptide | Kaa Light Chain Antibody Blocking Peptide |
| KDM1A Antibody Blocking Peptide | 组蛋白赖氨酸N-甲基ETMAR封闭多肽 |
| HHIPL1 Antibody Blocking Peptide | PACSIN2 Antibody Blocking Peptide |
| OIT3 Antibody Blocking Peptide | MAK10 Antibody Blocking Peptide |
| ADAM12 Antibody Blocking Peptide | RNF135 Antibody Blocking Peptide |
| SET Antibody Blocking Peptide | TMC8 Antibody Blocking Peptide |
| R3HCC1 Antibody Blocking Peptide | Recombinant Human MMP12 proteinPhospho-KSR1 (Ser392) Antibody Blocking Peptide |
| RASL11A Antibody Blocking Peptide | DNAJB13 Antibody Blocking Peptide |
| HABP2 27 kDa light chain Antibody Blocking Peptide | Phospho-LAT (Tyr171) Antibody Blocking Peptide |
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Recombinant Human MMP12 protein
￥1290 - 3960
