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Recombinant Human SLIT2 protei

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  • ¥1290 - 3960
  • 帛科
  • BK-DB8591
  • 国产
  • 2025年07月11日
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    • 详细信息
    • 文献和实验
    • 技术资料
    • 库存

      38

    • 英文名

      Recombinant Human SLIT2 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:
    货号 BK-DB8591 用途 仅供科研研究实验
    纯度 >90% as determined by SDS-PAGE 预测分子量 14.4 kDa
    表达宿主 E.coli 种属 Homo sapiens (Human)
    内毒素: Please contact with the lab for this information.
    蛋白构建: A DNA sequence encoding the human SLIT2 (Cys1416-Val1528) 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: Slit homolog 2 protein,Slit-2.
    背景介绍: Thought to act as molecular guidance cue in cellular migration, and function appears to be mediated by interaction with roundabout homolog receptors. During neural development involved in axonal navigation at the ventral midline of the neural tube and projection of axons to different regions. SLIT1 and SLIT2 seem to be essential for midline guidance in the forebrain by acting as repulsive signal preventing inappropriate midline crossing by axons projecting from the olfactory bulb. In spinal chord development may play a role in guiding commissural axons once they reached the floor plate by modulating the response to netrin. In vitro, silences the attractive effect of NTN1 but not its growth-stimulatory effect and silencing requires the formation of a ROBO1-DCC complex. May be implicated in spinal chord midline post-crossing axon repulsion. In vitro, only commissural axons that crossed the midline responded to SLIT2. In the developing visual system appears to function as repellent for retinal ganglion axons by providing a repulsion that directs these axons along their appropriate paths prior to, and after passage through, the optic chiasm. In vitro, collapses and repels retinal ganglion cell growth cones. Seems to play a role in branching and arborization of CNS sensory axons, and in neuronal cell migration. In vitro, Slit homolog 2 protein N-product, but not Slit homolog 2 protein C-product, repels olfactory bulb (OB) but not dorsal root ganglia (DRG) axons, induces OB growth cones collapse and induces branching of DRG axons. Seems to be involved in regulating leukocyte migration.
    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。

    产品细节图片1
    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。


    产品细节图片2
    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。

    产品细节图片3
    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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