IPTG is an analog of galactose that is non-metabolizable and inactivates the lac repressor to induce synthesis of β-galactosidase in E. coli. The expression of cloned genes under the control of the lac operon is induced by IPTG.It is also a substrate for thigalactosidetransacetylase and has been reported to induce penicillinase in bacteria.
IPTG is commonly used in cloning procedures that require induction of β-galactosidaseand is most often used with X-Gal (Gold Bio #X4281C) or Bluo-Gal (Gold Bio #B-673) for blue/white colony screening or Magenta-Gal (Gold Bio #B-378) for red/white colony screening of bacterial colonies.
IPTG is also used in the induction of recombinant proteins.In those systems, a protein of interest is encoded downstream of the IPTG inducible promoter.In the presence of IPTG, the protein of interest is induced in the cell culture.The culture can then be lysed and the protein expressed and purified through a number of methods, including His Tag or GST purification systems (for proteins with ligand tags).
GoldBio IPTG is made from a synthetic, non-animal origin.
Product Specifications:IPTGIsopropyl-beta-D-thiogalactoside
MOLECULAR BIOLOGY GRADE
Forumla: C9H18O5S
MW: 238.30 g/mol
Purity: >99% by HPLC
Dioxane free
Storage/Handling: Store desiccated at -20°C. Protect from light.
PubChem Chemical ID:656894
IPTG, which is similar to the lactose metabolite allolactose, induces the lac operon by binding to the lac repressor. Once IPTG binds to the lac repressor, a conformational change occurs, causing the lac repressor to dissociate from the lac promoter. Once the lac repressor dissociates, RNA polymerase binds to the promoter to initiate expression.
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Most papers recommend using between 0.1mM – 1.0 mM IPTG for expression. However, the amount you should use depends on your proteins. However, it is best to run an IPTG titration with ranges between 0.01mM – 1.0mM and compare using SDS-PAGE to determine the optimal concentration. Something to keep in mind is higher concentrations of IPTG can cause inclusion bodies.
IPTG can be dissolved in sterile water.
To make fresh 100mM IPTG stock solution:
Store powder IPTG desiccated at -20°C. Protect from light. For IPTG stock solution, store in 1 ml aliquots at -20°C for up to 1 year.
Primary applications of IPTG include:
IPTG, X-Gal and X-Gluc
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没错!最开始,Gold Biotechnology只生产这三个产品,30年后的今天,Goldbio的产品已经超过3000种,涵盖:
琼脂糖树脂
抗生素
生化试剂
缓冲液
克隆与诱导
培养基添加剂
洗涤剂
DNA蛋白质电泳
酶、抑制剂和底物
生长因子
植物研究
蛋白质表达与纯化
还原剂……
目前Goldbio最核心的产品是:荧光素钾盐,X-gluc, 双丙氨磷,草胺磷 和 X-α-gal