Expresso® Biotin C-His System (5 rxns)

• Protein binding studies
• Rapid protein purification
• Protein detection and ChIP Seq

Figure 1. Expresso Biotin vectors are available for C- or N-terminal biotinylation with optional SUMO tag to further enhance solubility and purification strategies.

• Seamless integration of your target protein with optimized expression elements
• No intervening restriction, protease or recombination sites
• Provides higher yields of perfect protein with no scars or unwanted cloning artifacts

	Lane 1 MW Ladder Lane 2 Uninduced cell lysate Lane 3 Induced cell lysate    Lane 4 Induced cell lysate, soluble fraction   Lane 5 Resin supernatant (unbound protein) Lane 6 Freed resin-bound target protein
Figure 2.  Expression and capture of biotinylated protein. Using a simple autoinduction method, a large amount of protein is expressed and is soluble (lanes 3-4). Due to efficient  in vivo biotinylation, most of the target protein is biotinylated and captured using biotin streptavidin purification (lanes 5-6).

Ultra-pure protein easier than ever

Expresso Biotin vectors are also available with the SUMO tag, which increases solubility, protein yield and provides a specific protease site. Using the SUMO N-Biotin version of the vector, you can bind your protein to streptavidin agarose and perform on-column cleavage, leaving you with stringently purified, native target protein.

Figure 3. Workflow diagram of AviTag capture and SUMO Protease on-column cleavage.

	Lane 1 MW Ladder Lane 2  Induced cell lysate (soluble fraction) Lane 3  Lysate after capture of biotinylated target   Lane 4  Streptavidin-captured protein Lane 5  Supernatant of untreated resin Lane 6  Streptavidin-captured protein, SUMO Protease treated Lane 7 Supernatant of resin, SUMO Protease treated
Figure 4. Expression and capture of biotinylated protein. A biotinylated protein is expressed and captured on streptavidin agarose (lanes 2-4). Without protease treatment, no protein is freed from the resin (lane 5). Streptavidin agarose was boiled to release all protein, which was then treated with protease to cleave the target (lane 6).

Four Expresso Biotin vector configurations for the ultimate flexibility in fusion tag applications:

Figure 5. Expressioneering Technology uses in vivo homologous recombination to seamlessly clone PCR-amplified DNA into the pAviTag  vectors. Target genes are amplified with primers that include 18 bases of overlap with the ends of the Expresso vector. The unpurified PCR amplicon is simply mixed with the pAviTag expression plasmids and immediately transformed into the highly competent cells provided. No DNA purification, enzyme treatment, vector preparation, or ligation steps are required

AviTag™ is a trademark of Avidity, LLC. AviTag™ technology (U.S. Patent Nos. 5,723,584, 5,874,239 & 5,932,433) is owned by and licensed from Avidity, LLC, Denver, Colorado.