大肠杆菌O157检测卡（食品安全检测）

广州健仑生物科技有限公司

广州健仑长期供应：动物、畜牧、食品、药品、化妆品、水产品、违禁品的快速检测试剂盒。

动物类的主要检测项目包括：猪、狗、猫、牛、鸡、鸭和鹅的传染病。

畜牧类的主要检测项目包括：多种添加剂、瘦肉精添加剂等。

食品类的主要检测项目包括：添加剂残留、农药残留、药物残留等。

化妆品的主要检测项目包括：重金属、有害物质等。

水产品的主要检测项目包括：呋喃类药物残留、抗生素残留等。

违禁品的主要检测项目包括：MOP/MET/KET/THC/MDMA/COC/BZO/AMP/K2/BAR/TCA/BUP/MTD/PCP/OXY/EDDP

我司还提供其它进口或国产试剂盒：包括传染病系列、免疫组化系列、诊断血清等产品欢迎致电。

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大肠杆菌O157检测卡（食品安全检测）

大肠杆菌O157检测卡（食品安全检测）

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【公司名称】 广州健仑生物科技有限公司

【市 场 部】 13802525278 020-82574011  杨永汉

【公司传真】 020-32206070

【腾讯Q Q】 2042552662

【公司地址】 广州清华科技园创新基地番禺石楼镇创启路63号二期2幢101-103室

Inclusion body contains mainly recombinant protein, but also contains some bacterial components, such as some outer membrane proteins, plasmid DNA and other impurities. Washing commonly used neutral detergent below 1%, such as Tween, Triton, Lubel and NP40 plus EDTA and reducing agent 2-mercapto-threitol (DTT), β-mercaptoethanol repeated multiple times, due to detergents The washing power increases with increasing ionic strength of the solution, plus 50 mM NaCL when washing inclusion bodies. The purity of inclusions extracted in this way can reach at least 50%, and can maintain the metathesis. Can also be used at low concentrations of guanidine hydrochloride or urea / neutral detergent / EDTA / reductant wash away the inclusion body surface adsorption of most of the insoluble protein. The pH of the washing liquid is similar to the physiological conditions of the engineering bacteria, and the reducing agent used is 0.1-5mM. EDTA is 0.1-0.3 mM. This step is important because detergents such as Triton X-100, deoxycholate and low concentrations of denaturant such as urea are thoroughly washed to remove impurities, since the E. coli outer membrane protein Omp T (37 KDa) is diluted in 4-8 mol / L urea Has proteolytic enzyme activity that can result in the degradation of recombinant proteins during inclusion and lysis.

For urea and guanidine hydrochloride choice:

Urea and guanidine hydrochloride are medium strength denaturants, easy to remove by dialysis and ultrafiltration. They have a strong reversible hydrogen bond denaturation of inclusion bodies, the required concentration of urea 8-10M, guanidine hydrochloride 6-8M. Urea dissolved inclusion body slower and weaker than the guanidine hydrochloride, the solubility of 70-90%, urea in the role of longer or at higher temperatures will be cracked to form cyanate, the amino group of the recombinant protein covalent modification, but dissolved in urea Has not ionization, neutral, low cost, protein renaturation will not cause a large number of protein precipitation and dissolution of inclusion bodies can be used for a variety of chromatographic purification, etc., it has been widely used.

Guanidine hydrochloride has a solubilization capacity of over 95%, and dissolves quickly without causing covalent modification of the recombinant protein. But it also has high cost, easy to produce precipitation under acidic conditions, the removal of refolding may cause a lot of protein precipitation and protein ion exchange chromatography interference shortcomings.

The final purification of inclusion bodies is generally ion exchange, hydrophobic, or affinity chromatography, affinity chromatography, especially metal chelate chromatography is more widely used, and the purification effect is not bad, usually one step to achieve purity 95% or more of inclusion bodies.