NZY Auto-Induction Kit (MB180)

49,00219,00

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Description: NZY Auto-Induction Kit is an innovative product developed to supplement your specific culture medium that allows E. coli to grow to high cell densities while promoting auto-induction of recombinant gene expression. NZY Auto-Induction Kit contains two concentrated sterile solutions and was designed for IPTG-inducible bacterial expression systems. Addition of these solutions to traditional complex media, such as Luria-Bertani (LB) broth, Terrific Broth (TB), results in high-level protein production. The kit simplifies induction of expression constructs by eliminating the need to take repeated OD readings and isopropyl β-D-1-thiogalactopyranoside (IPTG) addition. The method is based on medium components that are metabolized differentially to promote culture growth to high cell densities and subsequent induction of protein expression from lac-based promoters. NZY Auto-Induction Kit is ideal for high-throughput (HTP) methods, when you have to grow multiple cultures expressing various proteins simultaneously. NZYAI Solution 1 is the induction solution while it also contains magnesium and a mixture of trace metals. NZYAI Solution 2 is an optimized buffering solution.

Features:
– Self-induced protein expression system
– Flexibility: compatible to any conventional E. coli growth medium
– No need to monitor cell growth
– No need for IPTG induction

Applications:
– Used to supplement growth of E. coli cells transformed with a IPTG-inducible bacterial vector
– Ideal for high-throughput (HTP) methods, when you have to grow multiple cultures expressing different proteins simultaneously

High yields of recombinant proteins expressed using BL21(DE3) and BL21(DE3)pLysS E. coli strains as hosts
A range of experiments was performed to compare the efficiency of NZY Auto-Induction Kit and NZY Auto-Induction LB medium (powder). Both auto-induction systems were used for the production, in parallel, of 24 recombinant proteins from the anaerobic ruminal bacterium Ruminococcus flavefaciens in two expression E. coli strains: BL21(DE3) and BL21(DE3)pLysS. Recombinant proteins were purified through Immobilized Methal Affinity Chromatography (IMAC) and separated by SDS-PAGE. The data revealed that both auto-induction systems (kit or powder) performed at very high levels both in terms of cell density and recombinant protein expression.

 

NZY Auto-induction kit_figure1

Figure 1. Levels of purified proteins obtained from 24 different recombinant E. coli BL21(DE3) strains grown using NZY Auto-Induction Kit as supplement of LB media and NZY Auto-Induction LB medium (powder). The 24 recombinant proteins from Ruminococcus flavefaciens were purified through IMAC and levels of protein obtained evaluated (I) while degree of purification was confirmed through SDS-PAGE (II). M: Low Molecular Weight (LMW) Protein Marker.

 

NZY Auto-induction kit_figure2

Figure 2. Levels of purified protein obtained from 24 different recombinant E. coli BL21(DE3)pLysS strains grown using NZY Auto-Induction Kit as supplement of LB media and NZY Auto-Induction LB medium (powder). The 24 recombinant proteins from Ruminococcus flavefaciens were purified through IMAC and levels of protein obtained evaluated (I) while degree of purification was confirmed through SDS-PAGE (II). M: Low Molecular Weight (LMW) Protein Marker.

 

Storage: Store dry at room temperature. Once opened, store NZYAI Solution 1 at 4 ºC

Components:

– NZYAI Solution 1 (25 mL for 1 L or 5 x 25 mL for 5 L)

– NZYAI Solution 2 (50 mL for 1 L or 250 mL for 5 L)

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