Product Description
C3/FB-Depleted Serum is normal humanserum in which C3 and factor B(fB) have been removed by immunoaffinity chromatography.The product istested for the absence of C3 and factor B activity by testing classical and alternative pathway function and for the absence of C3 andfactorB proteins by double immunodiffusion. The C3/FB-depleted serumis certified to exhibitless than 5% classical pathway and alternativepathway activities. A functionalclassical pathway can be reconstitutedby addition of purified C3 protein (1.3mg/mL) whereas a functionalalternative pathway can be reconstituted byaddition of purified C3protein (1.3 mg/mL) and factor B protein (0.2 mg/mL)indicating thatall other complement components necessary for classical andalternativepathway activation are present. After reconstitution with C3 (1.3mgC3/mL) the C3/FB-Depleted Serum is certified to possess a functional classical pathway for complement activation whereas after reconstitutionwithC3 (1.3 mg C3/mL) and factor B (0.2 mg/mL), the product iscertified to possessa functional alternative pathway for complementactivation (Morgan, B.P.(2000); Dodds, A.W. and Sim, R.B. (1997)).Although a functional lectin pathwayshould also be reconstituted bythe addition of similar concentrations of C3,the function of thispathway is not tested.
Note that there is a weak C3by-passmechanism whereby the convertase C4b,C2a is able to activate C5(Rawal, N.& Pangburn, M.K. 2003) and form C5b-9 complexes capableof causing lowgrade lysis. This system of C5 activation is functionalin both the classicaland the lectin pathways and is approximately1000-fold less active than in thepresence of C3.
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谁能告诉我
不加磷酸化抑制剂也可以做磷酸化蛋白WB,但是有可能会影响具体的实验结果,大部分时候不加磷酸化抑制剂也能得到结果的。
需要注意蛋白的磷酸化是一个非常迅速的反应,所以取样品的过程要迅速,样品要新鲜。
所以做磷酸化蛋白WB,提取蛋白时不加磷酸酶抑制剂会有影响
(2)不同点:①影响因素:变构调节是由细胞内变构效应剂浓度的改变而影响酶的活性;化学修饰调节是激素等信息分子通过酶的作用而引起共价修饰。②酶分子改变:变构效应剂通过非共价键与酶的调节亚基或调节部位可逆结合,引起酶分子构像改变,常表现为变构酶亚基的聚合或解聚;化学修饰调节是酶蛋白的某些基团在其他酶的催化下发生共价修饰而改变酶活性。③特点及生理意义:变构调节的动力学特征为S型曲线,在反馈调节中可防止产物堆积和能源的浪费;化学修饰调节耗能少,作用快,有放大效应,是经济有效的调节方式。
相同点:都能以DNA为模板,从5'向3'进行核苷酸或脱氧核苷酸的聚合反应。
不同点
1、作用底物不同。RNA聚合酶底物是NTP;DNA聚合酶底物是dNTP。
2、RNA聚合酶作用不需要引物,而DNA聚合酶作用需要引物。
3、RNA聚合酶本身具有一定的解旋功能,而DNA聚合酶没有,当需要解开双链的时候要解旋酶和拓扑异构酶的帮助。
4、RNA聚合酶只具有5‘到3’端的聚合酶活性,而DNA聚合酶不仅有5‘到3’端的聚合酶活性,还具有3‘到5’端的外切酶活性。保证DNA复制时候校对,所以复制的忠实性高于转录的。
5、RNA聚合酶通常作用于转录过程;DNA聚合酶通常作用于DNA复制过程
protease
水解蛋白质肽键的一类酶的总称。按其水解多肽的方式,可以将其分为内肽酶和外肽酶两类。内肽酶将蛋白质分子内部切断,形成分子量较小的月示和胨。外肽酶从蛋白质分子的游离氨基或羧基的末端逐个将肽键水解,而游离出氨基酸,前者为氨基肽酶后者为羧基肽酶。按其活性中心和最适pH值,又可将蛋白酶分为丝氨酸蛋白酶、巯基蛋白酶、金属蛋白酶和酸性蛋白酶。按其反应的最适pH值,分为酸性蛋白酶、中性蛋白酶和碱性蛋白酶。工业生产上应用的蛋白酶,主要是内肽酶。
蛋白激酶
protein kinase
又称蛋白质磷酸化酶(protein phosphakinase)。一类催化蛋白质磷酸化反应的酶。

