Glutathione-Superflow allows rapid affinity purification of GST-tagged proteins for batch, gravity-flow, and FPLC applications. The resin is based on 6% cross-linked agarose, with glutathione covalently bound to the resin. It can withstand flow rates as high as 15ml/cm2/min and medium pressure (up to 150 psi).The GST Purification Kit contains sufficient buffers and Glutathione-Superflow Columns for performing five batch/gravity-flow purifications. Up to 10 mg per prep of GST-tagged recombinant proteins can be purified using this kit.
Glutathione-Superflow allows rapid affinity purification of GST-tagged proteins for batch, gravity-flow, and FPLC applications. The resin is based on 6% cross-linked agarose, with glutathione covalently bound to the resin. It can withstand flow rates as high as 15 ml/cm2/min and medium pressure (up to 150 psi).
The GST Purification Kit contains sufficient buffers and Glutathione-Superflow Columns for performing five batch/gravity-flow purifications. Up to 10 mg per prep of GST-tagged recombinant proteins can be purified using this kit.
GST tags
Epitope tags such as glutathione-S-transferase (GST) are often used as tags for proteins for expression and purification applications. Glutathione transferases are abundant enzymes involved in cellular defense against electrophilic chemical compounds, which bind glutathione with high affinity and specificity. The strength and selectivity of this interaction can be exploited to effectively purify GST-tagged proteins. Glutathione-Superflow Resin selectively binds the GST-tagged protein under normal conditions, allowing the protein of interest to be separated from whole-cell extracts rapidly and efficiently. A high degree of purification can be achieved in just one chromatographic step.
GST is a 35-kDa protein that can be assayed biochemically as well as immunologically. This characteristic sets it apart from many epitope tags that are simply short peptides. However, since GST is a large tag, the potential for degradation by proteases is higher than for smaller tags. Therefore, performing GST-based protein purification as quickly as possible under nondegrading conditions is necessary to minimize sample loss. GST loses its ability to bind Glutathione-Superflow Resin when denatured, and consequently strong denaturants such as guanidine-HCl or urea must not be used in the purification buffers. Check reagent compatibilities when designing your purification scheme.
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试剂盒里有详细的说明书,告诉你样品需要多少量,每个试剂需要加入多少量,和详细的实验步骤,一般买来就可以用,不用人教。
所以你问一个样需要多少量是没法回答的,测定过程是要加很多种试剂的。
1.模板提取(一般为RNA):Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定:分光光度计或NanoDrop
3.逆转录:逆转录试剂盒(或者一步法试剂盒),这一步可以用普通PCR做,也可以用水域做。
4.荧光定量PCR试剂:通常有用SYBR Green Mix做的,但是这里建议你用EvaGreen做,灵敏度和平行性都要好于SYBR Green,并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他:除了以上的那些还需要离心管、PCR管或板(Axygen反应比较好)、移液枪等,暂时就想到这么多。
EliKine™ 人 肝细胞生长因子 ELISA定量试剂盒
EliKine™ 人 干扰素-α ELISA定量试剂盒
EliKine™ 人 干扰素-γ ELISA定量试剂盒
EliKine™ 人 白介素-1α ELISA定量试剂盒
EliKine™ 小鼠 白介素-22 ELISA定量试剂盒
EliKine™ 小鼠 转化生长因子-β1 ELISA定量试剂盒
EliKine™ 小鼠 肿瘤坏死因子-α ELISA定量试剂盒
EliKine™ 小鼠 血管内皮生长因子 ELISA定量试剂盒
EliKine™ 大鼠 干扰素-γ ELISA定量试剂盒
EliKine™ 大鼠 白介素-1β ELISA定量试剂盒
EliKine™ 人 促甲状腺激素 ELISA定量试剂盒
只要引物设计的可以在基因组上做,那么理论上是没问题的!
只需要用RT-PCR荧光定量试剂盒里面的第二个,专门用来做Real-timePCR的那些试剂就可以了!

