Covalently bind proteins, peptides and other amine containing compounds.

Thermo Scientific Pierce Maleic Anhydride Activated Plates allow attachment of amine-containing peptides and other molecules to microplate wells for use in binding assays. The plates are useful for immobilizing peptides and other ligands that do not coat efficiently by passive adsorption. Reaction of the maleic anhydride groups with primary amines (-NH2) results in formation of amide bonds that are stable at neutral pH and above. Acidic conditions hydrolyze the bond; for example, the half-life of hydrolysis at pH 3.5 and 37°C is approximately 11 hours (Ref.1). Therefore, coating reactions are best performed at pH 8-9, and the reacted plates are best used at pH > 7 for ELISA and other methods.

Highlights:

• Spontaneously react with primary amines
• A direct, simple approach to covalently attach molecules to the well surface of an EIA plate
• Maleic anhydride maintains its integrity and availability for coupling for months under dry, room temperature conditions

Reaction scheme for coupling peptides and other amine-containing molecules.

Related Pierce Products

• Wash Buffers and additives:
• • BupH Modified Dulbecco’s PBS Packs (No. 28374)
  • BupH Tris-buffered Saline Packs (No. 28379)
  • Tween-20: Surfact-Amps 20 Purified Detergent Solution, 6 × 10 ml (No. 2832)

• Blocking Buffers:
  • SuperBlock Blocking Buffer in PBS (No. 37515) or TBS (No. 37535)
  • StartingBlock Blocking Buffer in PBS (No. 37538) or TBS (No. 37542)
  • Protein-Free Blocking Buffer in PBS (No. 37572) or TBS (No. 37570)

• Secondary Antibodies:
  • Many species: mouse, rabbit, goat, human, rat
  • Many forms: unconjugated, biotinylated, enzyme-labeled, and fluorophore-labeled

• Substrates:
  • 1-Step Ultra TMB (No. 34028)
  • QuantaBlu Fluorogenic Peroxidase Substrate Kit (No.15169)
  • SuperSignal ELISA Femto Substrate (No. 37075)

References:

1. Butler, P.J.G, et al. (1967). Reversible blocking of peptide amino groups by maleic anhydride. Biochem. J. 103, 78P
2. Dumoutier, L., et al. (2001). J. Immunol. 166, 7090-7095.
3. Marston, E.L., et al. (2002). Clin. Diagn. Lab. Immunol. 9, 446-452.
4. Leu, S.-J., et al. (2003). J. Biol. Chem. 278, 33801-33808.
5. Liao, Y.-F., et al. (2002). J. Biol. Chem. 277, 14467-14474.
6. Batista, F.D. and Neuberger, M.S. (2000). EMBO J. 19, 513-520.