Capture His-tagged fusion proteins for easy plate-based assays.

Thermo Scientific Pierce Nickel Coated Plates provide a simple format for protein:protein interaction studies. These Ni(2+) chelate-coated plates are ideal for analyzing polyhistidine-tagged fusion proteins by ELISA-based methods. Proteins that contain a succession of several histidine residues at the amino or carboxyl terminus have a strong binding affinity for metal. Bacterial lysates containing polyhistidine-tagged fusion proteins can be added directly to the plates without the need for blocking. The clear, white or black plates can be used with colorimetric, chemiluminescent or fluorescent detection methods, respectively.

Highlights:

• Ni²⁺ activated surface enables metal-chelate binding of His-tagged proteins
• Detergents used to lyse cells don't inhibit binding to activated plates as they do with plain polystyrene
• Better binding for sensitive assays compared to other commercially available nickel-activated plates
• Detection limit: 1 ng of polyhistidine fusion protein
• Binding capacity: ~9 pmol His-tagged protein (27 kDa)/well
• Activation level: 200 µl

Binding performance of nickel coated plates. Comparison of Thermo Scientific Pierce Nickel Coated Plates (Pierce) to Nickel-chelate plates from Brand Q in binding performance for His-tagged ATF fusion protein. Detection was accomplished with an HRP-conjugated anti-ATF antibody and TMB substrate.

References:

1. Desai, S., et al. (1997). Previews 1(4), 12-15.
2. Tu, Y., et al. (1999). Mol. Cell. Biol. 19, 2425-2434.
3. Pan, W., et al. (2003). J. Biol. Chem. 278, 27820-27827.