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MarkerGeneTM Glucuronidase (GUS) Reporter GeneActivity Detection Kit
背景描述:
報告基因普遍用做“標志物 Markers”對基因調控和定位分析,以及突變基因分析。通過免疫實驗,生化活性或組織化學分析來對組織或細胞的報告基因表達進行檢測。
大腸桿菌來源的β-葡萄糖苷酶(β-glucuronidase,GUS)普遍認為是轉基因植物研究中非常理想的報告基因(marker gene)。GUS報告基因系統(tǒng)具有許多優(yōu)勢,包括穩(wěn)定表達大腸桿菌GUS,不會干擾正常植物代謝和高等植物內幾乎無內源性GUS活性。
數種葡萄糖醛酸底物可用來檢測GUS表達,這些底物都包含糖基D-葡萄糖醛酸苷(D-glucopyranosiduronic acid)通過糖苷鍵連接到生色底物、熒光底物或其他檢測分子的—OH。體外用于GUS活性檢測的最廣泛使用的熒光底物為MUG(4-Methylumbelliferyl beta-D-glucuronide,4-甲基傘型酮-beta-D-葡糖苷酸,CAS:6160-80-1或881005-91-0)。一旦被水解,生成熒光素4-MU(4-甲基傘型酮)和β-D-葡萄糖醛酸。
檢測原理:
本MarkerGeneTM GUS報告基因活性檢測試劑盒(MarkerGeneTM Glucuronidase(GUS) Reporter Gene Activity Detection Kit)以MUG為熒光顯色底物。當植物或其他細胞用GUS萃取液(含磷酸-EDTA,pH 7.0和去污劑)萃取后,所得到的萃取液(含GUS)能水解4-MUG生成4-MU(4-甲基傘型酮,pKa 8.2)和β-D-葡萄糖醛酸。加入碳酸鈉緩沖液終止該反應(因4-MU在pH值高于pKa的情況下發(fā)射最大熒光信號)。4-MU的最大激發(fā)波長為365nm,最大發(fā)射波長為455nm。
試劑盒組分:
組分名稱 |
組分編號 |
規(guī)格 |
保存 |
GUS Extraction Buffer |
0877-001 |
25 mL |
Store cold (4℃) |
Carbonate Stop Buffer |
0877-002 |
25 mL |
Store cold (4℃) |
GUS Assay Buffer |
0877-003 |
5 mL |
Store cold (4℃) |
4-MU calibration stock solution |
0877-004 |
5 mL |
Store cold (4℃) |
訂購信息:
貨號 |
產品名稱 |
規(guī)格 |
價格(元) |
M0877 |
MarkerGene? β-Glucuronidase (GUS) Reporter Gene Activity Detection Kit |
1Kit |
5680.00 |
使用文獻(MarkerGene? β-Glucuronidase(GUS) Reporter Gene Activity Detection Kit):
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