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- 详细信息
- 技术资料
- 库存:
44
- 英文名:
Recombinant Human S100A16 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
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 S100A16 (Ser2-Ser103) 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: AAG13 Protein, Human; DT1P1A7 Protein, Human; MGC17528 Protein, Human; S100F Protein, Human
背景介绍: S1A16 is a member of S1 protein super family that carries calcium-binding EF-hand motifs. S1 proteins are cell- and tissue-specific and are involved in many intra- and extracellular processes through interacting with specific target proteins. S1A16 expression was found to be astrocyte-specific. The S1A16 protein was found to accumulate within nucleoli and to translocate to the cytoplasm in response to Ca(2+) stimulation. The homodimeric structure of human S1A16 in the apo state has been obtained both in the solid state and in solution, resulting in good agreement between the structures with the exception of two loop regions. The homodimeric solution structure of human S1A16 was also calculated in the calcium(II)-bound form. Differently from most S1 proteins, the conformational rearrangement upon calcium binding is minor. Immunoprecipitation analysis revealed that S1A16 could physically interact with tumor suppressor protein p53, also a known inhibitor of adipogenesis. Overexpression or RNA interference-initiated reduction of S1A16 led to the inhibition or activation of the expression of p53-responsive genes, respectively. S1A16 protein is a novel adipogenesis-promoting factor.
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:
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。
| 货号 | BK-DB4377 | 用途 | 仅供科研研究实验 |
| 纯度 | >90% as determined by SDS-PAGE | 预测分子量 | 12.92 kDa |
| 表达宿主 | E.coli | 种属 | Homo sapiens (Human) |
蛋白构建: A DNA sequence encoding the human S100A16 (Ser2-Ser103) 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: AAG13 Protein, Human; DT1P1A7 Protein, Human; MGC17528 Protein, Human; S100F Protein, Human
背景介绍: S1A16 is a member of S1 protein super family that carries calcium-binding EF-hand motifs. S1 proteins are cell- and tissue-specific and are involved in many intra- and extracellular processes through interacting with specific target proteins. S1A16 expression was found to be astrocyte-specific. The S1A16 protein was found to accumulate within nucleoli and to translocate to the cytoplasm in response to Ca(2+) stimulation. The homodimeric structure of human S1A16 in the apo state has been obtained both in the solid state and in solution, resulting in good agreement between the structures with the exception of two loop regions. The homodimeric solution structure of human S1A16 was also calculated in the calcium(II)-bound form. Differently from most S1 proteins, the conformational rearrangement upon calcium binding is minor. Immunoprecipitation analysis revealed that S1A16 could physically interact with tumor suppressor protein p53, also a known inhibitor of adipogenesis. Overexpression or RNA interference-initiated reduction of S1A16 led to the inhibition or activation of the expression of p53-responsive genes, respectively. S1A16 protein is a novel adipogenesis-promoting factor.
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 APE1 | Recombinant human Myoglobin, His |
| 重组人TNMD蛋白 | NKX1-1 Antibody Blocking Peptide |
| 磷酸化载脂蛋白A1封闭多肽 | PBX1 Antibody Blocking Peptide |
| 转录因子OTX1+OTX2封闭多肽 | Phospho-NCOA3 (Thr24) Antibody Blocking Peptide |
| 重组人Toll样受体2蛋白 | AKR1A1 Antibody Blocking Peptide |
| 跨膜蛋白208封闭多肽 | phospho-c-ABL (Tyr245) isoform b (1149aa) Antibody Blocking Peptide |
| 角蛋白相关蛋白KAP27.1封闭多肽 | CK II beta Antibody Blocking Peptide |
| ZCWPW2蛋白封闭多肽 | 磺胺嘧啶 |
| 转录因子相关蛋白NK1封闭多肽 | ZCWPW2 Antibody Blocking Peptide |
| B淋巴细胞白血病前体蛋白转录因子1封闭多肽 | KAP27.1 Antibody Blocking Peptide |
| NADP依赖的乙醇脱氢酶封闭多肽 | TMEM208 Antibody Blocking Peptide |
| 磷酸化类固醇受体辅助活化因子-3 | Recombinant human TLR2 protein, His |
| 磷酸化非受体酪氨酸激酶c-Abl封闭多肽 | Recombinant Human S100A16 proteinOTX1 + OTX2 Antibody Blocking Peptide |
| 丝/苏氨酸蛋白激酶II β封闭多肽 | phospho-FAS (Tyr291) Antibody Blocking Peptide |
| 重组人肌红蛋白 | Recombinant human TNMD protein, His |
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。
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Recombinant Human S100A16 protein
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