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Cathepsin B His tag Protein, Human

APP secretase (APPS),Cathepsin B1,CPSB

价格 ￥1,850.00 （ 1-2周）
货号 UA100041
规格
数量
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产品介绍

产品介绍 评论(0)

产品规格

  • 物种

    Human

  • 分子别名

    APP secretase (APPS), Cathepsin B1, CPSB

  • Accession

    P07858

  • 表达序列

    Met1-Ile339, with C-10*His

  • 表达宿主

    HEK293

  • 分子量

    Predicted MW: 39.5 kDa Observed MW: 43 kDa

  • 纯度

    >95% by SDS-PAGE

  • 内毒素含量

    <1EU/μg

  • 活性

    Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC (Catalog # ES008). The specific activity is >10000 pmol/min/μg, as measured under the described conditions.

  • 标记

    Unconjugated

  • 标签

    His Tag

  • 性状

    Lyophilized Powder

  • 缓冲体系

    0.2M PBS, pH7.4.

  • 溶解方法

    Reconstitute no more than 1 mg/mL according to the size in deionized water after rapid centrifugation.

  • 储存条件


    12 months from date of receipt, -20 to -70 °C as supplied.
    6 months, -20 to -70 °C under sterile conditions after reconstitution.
    1 week, 2 to 8 °C under sterile conditions after reconstitution.
    Please avoid repeated freeze-thaw cycles.

背景介绍

  • Cathepsin B belongs to a family of lysosomal cysteine proteases known as the cysteine cathepsins and plays an important role in intracellular proteolysis. In humans, cathepsin B is encoded by the CTSB gene. Cathepsin B is upregulated in certain cancers, in pre-malignant lesions, and in various other pathological conditions. Cathepsin B may enhance the activity of other proteases, including matrix metalloproteinase, urokinase (serine protease urokinase plasminogen activator), and cathepsin D, and thus it has an essential position for the proteolysis of extracellular matrix components, intercellular communication disruption, and reduced protease inhibitor expression. Cells may become carcinogenic when cathepsin B is unregulated. Cathepsin B has been proposed as a potentially effective biomarker for a variety of cancers. Overexpression of cathepsin B is correlated with invasive and metastatic cancers.

操作步骤

  • 实验方法
    实验原理：检测Cathepsin B His Tag Protein, Human切割荧光肽底物Z-LR-AMC的能力。
    实验材料
    1.蛋白激活缓冲液：25 mM MES, 5 mM DTT, pH 5.0
    2.检测缓冲液：25 mM MES,0.02% Brij-35, pH 5.0
    3.Cathepsin B His Tag Protein, Human
    4.底物：Z-LR-AMC (ES008)
    5.96 ELISA Removable Plate, Black, High binding (GENEVER, Catalog # GMO2-96H)
    6.读板器（PerkinElmer，激发波长380 nm，发射波长460 nm）
    实验步骤
    1.Cathepsin B His Tag Protein, Human (10 μg/mL) 在蛋白激活缓冲液中于室温下激活15分钟，然后进行反应。
    2.在检测缓冲液中将激活的Cathepsin B His Tag Protein, Human稀释至0.4 µg/mL、0.2 µg/mL、0.1 µg/mL、0 µg/mL。
    3.在检测缓冲液中将Z-LR-AMC稀释至80 µM。
    4.向黑色孔板中加入稀释后的Cathepsin B His Tag Protein, Human，每孔50 µL，然后加入50 µL的80 µM底物。
    5.在激发和发射波长分别为380 nm和460 nm的情况下以动力学模式读取荧光信号5分钟。
    6.计算特异性活性：

    Specific Activity (pmol/min/µg) =

    Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)

    amount of enzyme (µg)

    *底物空白校正
    **使用校准标准品7-氨基-4-甲基香豆素（7-amino-4-Methyl Coumarin）推导

  • 电泳

    • 2μg（R: reducing conditions）

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