QIAexpress Type IV Kit

For high-level expression and one-step purification of N-terminally His-tagged proteins

Vectors covering all three reading frames
Complete kit for expression and purification of His-tagged proteins
High-level expression of N-terminally His-tagged proteins
Versatile, complete system for one-step purification and sensitive detection

The QIAexpress System provides materials for expression, purification, detection, and assay of His-tagged proteins.

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Product Details
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Product		Cat. no.	List price:
QIAexpress Type IV Kit 5 μg each: pQE-30, pQE-31, pQE-32 (N-terminal 6xHis); 10 ml Ni-NTA Agarose		32149	$930.00	Add to cart

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The QIAexpress Type IV Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Performance

The QIAexpress Type IV Kit gives high expression, up to 50% of total cellular protein.

Principle

QIAexpress pQE vectors combine a powerful phage T5 promoter (recognized by E. coli RNA polymerase) with a double lac operator repression module to provide tightly regulated, high-level expression of recombinant proteins in E. coli. Protein synthesis is effectively blocked in the presence of high levels of lac repressor and the stability of cytotoxic constructs is enhanced. The pQE vectors (see figure pQE Vectors and table) enable placement of the His tag at either the N- or C-terminus of the recombinant protein.

Elements present in QIAexpress pQE Vectors
Element	Description
1. Optimized promoter/operator element	Consists of the phage T5 promoter and two lac operator sequences, which increase the probability of lac repressor binding and ensure efficient repression of the powerful T5 promoter
2. Synthetic ribosomal binding site RBSII	For efficient translation
3. His-tag coding sequence	Either 5' or 3' to the polylinker cloning region
4. Translational stop codons	In all reading frames for convenient preparation of expression constructs
5. Two strong transcriptional terminators	t0 from phage lambda, and T1 from the rrnB operon of E. coli, to prevent read-through transcription and ensure stability of the expression construct
6. ColE1 origin of replication	From pBR322
7. beta-lactamase gene (bla)	Confers ampicillin resistance

Procedure

Inserts encoding proteins of interest are cloned into appropriate constructs and transformed into a suitable E. coli strain for expression. Expression is induced by addition of IPTG. QIAexpress Type IV Kit constructs can be transformed into E. coli, used as a shuttle vector for recombinant protein expression in insect cells, or transfected into mammalian cells.

Applications

The QIAexpress Expression System provides high-level expression of proteins suitable for many applications, including:

Purification of functional, conformationally active proteins
Purification under denaturing conditions for antibody production
Crystallization for determination of three-dimensional structure
Assays involving protein-protein and protein-DNA interactions

FAQs

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	What are the most commonly used protease inhibitors? FAQ ID -53	View
	How can I increase the amount of soluble recombinant protein in E. coli expression? FAQ ID -64	View
	How can I remove imidazole from a protein sample? FAQ ID -91	View
	How can I eliminate contaminating protein in my Ni-NTA 6xHis-tag protein purification? FAQ ID -102	View
	How does imidazole affect my quantitation of protein? FAQ ID -132	View
	How do I prepare an insert for pQE vectors? FAQ ID -185	View
	What are the features and benefits of the QIAexpress 6xHis Tag System? FAQ ID -193	View
	Can I use HEPES buffer instead of phosphate in my Ni-NTA column? FAQ ID -291	View
	What is the origin of replication and the plasmid copy number of the pQE vectors? FAQ ID -338	View
	How can I improve the expression of proteins containing hydrophobic regions? FAQ ID -339	View
	Which primers can I use for sequencing pQE-expression vector constructs? FAQ ID -343	View
	What is the composition of PBS? FAQ ID -361	View
	Why do you recommend using Triton X for the purification of 6xHis-tagged protein? -100	View
	What is the difference between Ni-NTA Agarose and Ni-NTA Superflow? FAQ ID -764	View
	How can I be sure that I am harvesting my induced bacterial culture at the best time point for protein expression? FAQ ID -788	View
	Are the buffers in the Ni-NTA Fast Start Kit the same as the ones for use with Ni-NTA purchased separately? FAQ ID -791	View
	How can I increase expression of my 6xHis-tagged protein in E. coli? FAQ ID -63	View

Kit Handbooks

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	The QIAexpressionist - (EN) A handbook for high-level expression and purification of 6xHis-tagged proteins	Show details

Supplementary Protocols

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	QIAexpress® Data Sheet: E. coli strain M15 - (EN)	Show details
	QIAexpress® Data Sheet: Genotype analysis of E. coli strains SG13009 and M15 - (EN)	Show details

Quick-Start Protocols

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	Automated Purification of 6xHis-tagged Proteins from E. coli Using Ni-NTA Superflow under Native Conditions (EN)	Show details
	Ni-NTA Agarose Purification of 6xHis-tagged Proteins from E. coli under Native Conditions (EN)	Show details

Selection Guides

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	Overview of QIAGEN’s pQE Vectors	Show details

Safety Data Sheets

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	MSDS QIAexpress Type IV Kit

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