Fig1. IPTG induction of GST-fusion protein production in BL21 E. coli.
Figure1. IPTG induction of GST-fusion protein production in BL21 E. coli. IPTG at 0.5 mM from Hello Bio was used to drive expression of a GST fusion protein under the control of the lac operon. This is visualised on a Coomassie stained PVDF membrane where protein expression was first induced then purified using a glutathione column (W1-W4: washes, E1-E3: Elution fractions with reduced glutathione). For more details see #Protocol 1 in application notes below.
Fig2. IPTG induction of GST production in BL21 E. coli.
Figure2. IPTG induction of GST production in BL21 E. coli. IPTG at 0.5 mM from Hello Bio was used to drive expression of GST under the control of the lac operon. This is visualised on a Coomassie stained PVDF membrane where protein expression was first induced then purified using a glutathione column (W1-W4: washes, E1-E3: Elution fractions with reduced glutathione). For more details see #Protocol 1 in application notes below.
Fig1. IPTG induction of GST-fusion protein production in BL21 E. coli.
Figure1. IPTG induction of GST-fusion protein production in BL21 E. coli. IPTG at 0.5 mM from Hello Bio was used to drive expression of a GST fusion protein under the control of the lac operon. This is visualised on a Coomassie stained PVDF membrane where protein expression was first induced then purified using a glutathione column (W1-W4: washes, E1-E3: Elution fractions with reduced glutathione). For more details see #Protocol 1 in application notes below.
Fig2. IPTG induction of GST production in BL21 E. coli.
Figure2. IPTG induction of GST production in BL21 E. coli. IPTG at 0.5 mM from Hello Bio was used to drive expression of GST under the control of the lac operon. This is visualised on a Coomassie stained PVDF membrane where protein expression was first induced then purified using a glutathione column (W1-W4: washes, E1-E3: Elution fractions with reduced glutathione). For more details see #Protocol 1 in application notes below.
Biological Data
Biological description
As an allolactose mimic, IPTG can induce the lac operon and is therefore commonly used to induce protein expression.
IPTG is commonly used with X-Gal in blue/white screening for colony selection.
Application notes
#Protocol 1: IPTG induction of protein production in E. coli
BL21 E. coli were transformed with plasmids containing GST or a GST fusion protein under the control of the lac operon using standard protocols (see Froger and Hall., 2007).
Transformed colonies were cultured in 2xYT medium containing 50µg/ml kanamycin (HB4429) at 10ml scale before this was transferred into 1L cultures and cultured at 37°C.
Cells were cultured until OD600 reached 0.6 before protein expression was induced with the adittion of 0.5mM IPTG and the temperature was reduced to 25ºC.
Cells were grown for a further 4 hours before being spun at 6,400g for 15 minutes at 4°C to pellet cells. This was then resuspended in lysis buffer (25mM Tris, 150mM NaCl, 10% glycerol, 1% triton X-100, 1x protease inhibitors (HB9081), pH7.5), sonicated and allowed to solubilise before then being spun at 48,300g for 25 minutes at 4°C.
The supernatent was removed and subsequently purified using glutathione beads.
Samples from all stages of the protein production process were loaded onto a 15% acrylamide gel, run at 90/170V then transferred to PVDF (400mA/90 mins) before being visualised using Coomassie dye (HB0739).
Solubility & Handling
Storage instructions
+4°C
Solubility overview
Soluble in water (100 mM)
Important
This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use.
![IPTG [367-93-1] Chemical Structure](https://hellobio.com/media/catalog/product//h/b/hb3941.png "IPTG [367-93-1]")
Understanding purity and quality - a guide for life scientists
