Successfully coat sulfhydryl peptides in 96-well microplates.

Thermo Scientific Pierce Maleimide Activated Plates are ideal for binding sulfhydryl-containing molecules that are difficult to coat onto polystyrene plates, such as peptides that contain a terminal cysteine. These coated plates are an especially useful tool for assessing specific anti-hapten antibody titers during antibody production. Maleimide Activated Plates are available in clear for colorimetric assays, white for chemiluminescent assays, and black for fluorescent assays.

Highlights:

• Pre-blocked to reduce nonspecific binding
• Convenient 8-well strip format
• Easy (spontaneous) immobilization of peptides derivatized with a terminal cysteine and proteins with free sulfhydryl
• Binding Capacity: 100-150 pmol of a sulfhydryl-containing peptide (307 Da)/well
• Activation Level: 100 µl

Reaction scheme for coupling peptides and other sulfhydryl-containing molecules

Important Product Information

Maleimides react with free sulfhydryl group(s) at pH of 6.5-7.5, forming stable thioether linkages. Reaction with amines becomes significant at pH > 7.5. Some sulfhydryl-containing peptides and proteins may oxidize in solution and form disulfide bonds, which cannot react with maleimides. Disulfide bonds can be reduced to produce free sulfhydryls. The TCEP Disulfide Reducing Gel enables peptide or protein reduction while recovering the sample in the absence of reducing agents. Sulfhydryls can be introduced via amine modification using N-succinimidyl S-acetylthioacetate (SATA) or 2-iminothiolane•HCl (Traut’s Reagent).

References:

1. Ostrowski, M., et al. (2002). J. Virol. 76, 4241-4250.
2. Nisitani, S., et al. (1999). Proc. Natl. Acad. Sci. USA 96, 2221-2226.
3. Reinhard, M., et al. (1999). J. Biol. Chem. 274, 13410-13418.