MAT-Tag™ Protein Expression System

Combine IMAC Versatility with the Sensitivity of FLAG® Detection

The MAT-Tag™ System (metal affinity tag) utilizes a small, novel, seven amino acid (HNHRHKH) sequence created for IMAC purification of recombinant MAT fusion proteins and the high sensitivity detection of the antibody based FLAG^® System.

• Dual tagged fusion proteins
• N- or C- terminal tagging configurations
• Compatible to Nickel and Cobalt affinity resins, such as HIS-Select™
• Superior antibody detection

Data

Technical Posters

• The MAT™ Tag System*: A Novel Histidine-based Metal Affinity Tag (MAT) System for Expression, Purification and Detection of Recombinant Fusion Proteins
• Expression, Purification and Detection of Recombinant Fusion Proteins Using the MAT™ Tag System

MAT Bacterial Expression Vectors
tac Promoter System
T7 Promoter System
T7 Promoter System with Enterokinase Recognition Site

MAT Mammalian Expression Vectors
Transient Expression

MAT Baculovirus Transfer Vectors

MAT Monoclonal Antibody

FLAG Antibody

Data

Figure 1. Full-length FLAG-BAP-MAT fusion protein is detected on Western blots by monoclonal antbodies to the FLAG and MAT tags. Lysates from uninduced (lanes 2 and 5) and induced (lanes 3 and 6) E. coli cultures and ColorBurst Markers (lanes 1 and 4) were seperated by SDS-PAGE and blotted to nitrocellulose. The resulting blot was immunostained using either ANTI-FLAG® M2-HRP conjugate (lanes 1-3) or Anti-MAT monoclonal antibody (0.5 µg/ml) followed by Rabbit-Anti-Mouse IgG-HRP conjugate (lanes 4-6). The blots were developed and visualized with TMB Substrate.

Figure 2. MAT-tagged fusion protein expressed in E. coli can be affinity purified by IMAC. The coding region for GrpE was cloned into an E. coli expression plasimd, pFLAG-MAC, with an N-terminal FLAG tag and a C-terminal MAT tag. The fusion protein was expressed in E. coli strain BL21. A lysate (L, lane 2) was made in CelLytic B and separated by centrifugation into a pellet (P, lane 3) and a supernatant (S, lane 4) fraction. The supernatant was applied to a 1.0 mL HIS-Select HF Nickel Affinity Gel column and the flow through (F, lane 5), wash (1-3, lanes 6-8) and elution (1-4, lanes 9-12) fractions were collected. Samples of the fractions and ColorBurst Markers (lane 1) were seperated by SDS-PAGE and visualized by EZBlue staining.

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MAT Bacterial Expression Vectors

tac Promoter System

Vectors utilizing the strong tac promoter (a hybrid of the E. coli trp and lac promoters) offer protein expression levels in excess of 10 mg/L of culture when using IPTG as a de-repressor. These vectors can be used to express protein in any established E. coli host.

pTAC-MAT™-1 Cytoplasmic expression of N-terminal MAT fusion proteins under the tac promoter. Supplied with a pTAC-MAT-2+BAP Control Vector. Product # Product Name Size E5530 pTAC-MAT-1 Expression Vector 10 µg
pTAC-MAT™-2 Cytoplasmic expression of C-terminal MAT fusion proteins under the tac promoter. Supplied with a pTAC-MAT-2+BAP Control Vector. Product # Product Name Size E5405 pTAC-MAT-2 Expression Vector 10 µg

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MAT Bacterial Expression Vectors

T7 Promoter System

The pT7-MAT™ vectors offer the very strong T7/lac promoter. These expression vectors produce even higher yields of recombinant proteins than the tac promoter system. However, the T7 promoter is known for background (“leaky”) expression, which can be a drawback when recombinant proteins are toxic to the host cell. Therefore, Sigma’s vectors contain the lac operator (lacO) sequences immediately downstream from the promoter to reduce leaky expression. Unlike the tac promoter system, pT7 vectors must be expressed in hosts containing a source of the T7 polymerase such as (DE3) lysogenic strains.

	pT7-MAT™-1 Cytoplasmic expression of N-terminal MAT fusion proteins under the T7llac promoter. Supplied with a pT7-MAT-2+BAP Control Vector. Product # Product Name Size E5780 pT7-MAT-1 Expression Vector 10 µg
	pT7-MAT™-2 Cytoplasmic expression of C-terminal MAT fusion proteins under the T7llac promoter. Supplied with a pT7-MAT-2+BAP Control Vector. Product # Product Name Size E5655 pT7-MAT-2 Expression Vector 10 µg
	pT7-FLAG-MAT™-1 Cytoplasmic expression of N-terminal Met-FLAG, C-terminal MAT dual tagged fusion proteins under the T7llac promoter. Supplied with a pT7-FLAG-MAT-1+BAP Control Vector. Product # Product Name Size E5280 pT7-FLAG-MAT-1 Expression Vector 10 µg
	pT7-MAT-FLAG™-2 Cytoplasmic expression of N-terminal MAT, C-terminal FLAG dual tagged fusion proteins under the T7llac promoter. Supplied with a pT7-FLAG-MAT-1+BAP Control Vector. Product # Product Name Size E4905 pT7-MAT-FLAG-2 Expression Vector 10 µg

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MAT Bacterial Expression Vectors

T7 Promoter System with Enterokinase Recognition Site

The recognition sequence for enterokinase, Asp-Asp-Asp-Asp-Lys, is found at the C-terminal end of the FLAG^® epitope tag. Removal of the FLAG is possible in all fusion proteins containing an N-terminal FLAG sequence. Dual tag fusion proteins may also be cleaved with enterokinase for removal of one or more tags, depending on the position of FLAG in the protein sequence.

pT7-MAT-FLAG™-1 Cytoplasmic expression of N-terminal MAT-FLAG dual tagged fusion proteins under the T7llac promoter. Supplied with a pT7-FLAG-MAT-1+BAP Control Vector. Product # Product Name Size E5155 pT7-MAT-FLAG-1 Expression Vector 10 µg
pT7-FLAG-MAT™-2 Cytoplasmic expression of C-terminal FLAG-MAT dual tagged fusion proteins under the T7llac promoter. Supplied with a pT7-FLAG-MAT-1+BAP Control Vector. Product # Product Name Size E5030 pT7-FLAG-MAT-2 Expression Vector 10 µg

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MAT Mammalian Expression Vectors
Transient Expression

CMV vectors contain the pMB1 (derivative of pBR322) origin for replication in bacterial cells, the β-lactamase gene for ampicillin resistance selection in bacteria, hGH, polyA, and the f1 origin.

pCMV-FLAG-MAT™-1 Transient cytoplasmic expression of N-terminal MeT-FLAG, C-terminal MAT dual tagged fusion proteins under the CMV promoter. Supplied with pCMV-FLAG-MAT-1 MAPK1 Control Vector. Product # Product Name Size C5864 pCMV-FLAG-MAT-1 Expression Vector 20 µg
pCMV-MAT-FLAG™-1 Transient cytoplasmic expression of N-terminal MaT-FLAG dual tagged fusion proteins under the CMV promoter. Supplied with pCMV-FLAG-MAT-1 MAPK1 Control Vector. Product # Product Name Size C5989 pCMV-MAT-FLAG-1 Expression Vector 20 µg
pCMV-FLAG-MAT™-2 Transient cytoplasmic expression of C-terminal FLAG-MAT dual tagged fusion proteins under the CMV promoter. Supplied with pCMV-FLAG-MAT-1 MAPK1 Control Vector. Product # Product Name Size C6114 pCMV-FLAG-MAT-2 Expression Vector 20 µg

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MAT Baculovirus Transfer Vectors

pPolh-MAT™ are baculovirus transfer vectors used for producing Metal Affinity Tag (MAT™-Tag) fusion proteins in insect cells. The pPolh-MAT vectors contain the strong viral polyhedrin (polh) promoter for high-level expression of target genes during the very late phase of infection. The vectors also contain a high copy of bacterial origin of replication and an ampicillin resistance gene (amp^r) for easy propagation in E. coli host cells.

MAT Antibody

FLAG Antibody

For more information on our FLAG product offering, visit the FLAG System: Recombinant Protein Purification, Detection and Expression page.

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