The use of peptides for the generation of antibodies against specific peptides has become an essential tool in proteomic research.
A crucial step in using peptides as antigens is their coupling to carrier proteins. Peptides are designed to be good epitopes by themselves; however they fail to elicit a strong antibody response. The reason for this is that the epitopes are able to bind to the B-cells, but are too small to contain additional motifs required to associate with class II molecules and T-cell receptors. The carrier proteins, once coupled to the peptides, provide these additional motifs and generate a strong antibody response.
The HOOK™ Peptide Coupling kits allow researchers to attach peptides to a carrier protein of choice using either primary amines or sulfhydryls in their peptide. The HOOK™ Peptide Coupling kits are supplied with one of our carrier proteins in convenient single use OneQuant™ format or without a carrier protein. Both kits can be supplied with or without carrier proteins.
The carrier proteins are bovine serum albumin (BSA), keyhole limpet hemocyanin (KLH) and HyperCarrier™, a cationized BSA. For further details on the carrier protein properties see the carrier protein section.
HOOK™ Peptide Coupling Kit (Amine Reactive)
This kit is designed for the coupling of peptides to carrier proteins, utilizing the primary amines and carboxyl groups of the peptides and carrier proteins. This kit utilizes the chemical heterobifunctional crosslinker EDC (1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride) to couple peptides to carrier proteins. EDC first reacts with the carboxyl groups, producing an amine reactive intermediate, O-acylisourea that rapidly reacts with the amine groups of the peptide/protein.
HOOK™ Peptide Coupling Kit (Sulhydryl Reactive)
A common practice in the design of peptides is the addition of a cysteine residue at the N- or C-terminus of the peptide, which can be used for coupling to carrier proteins. This kit is designed for the coupling of peptides to carrier proteins, utilizing a sulfhydryl group in the peptide.
This kit exploits the chemical cross-linker sulfoSMCC (Sulfosuccinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate) to couple peptides through their sulfhydryl groups, found on cysteine side chains, to the primary amines on the carrier proteins.
The N-hydroxysuccinimide (NHS) ester in sulfoSMCC reacts with primary amines to form covalent amide bonds and the maleimide group reacts with sulfhydryl groups to form stable thioether bonds.
If peptides do not contain a sulfhydryl group then G-Biosciences recommends and supplies Traut’s reagent (2-IminothiolaneHCl). Traut’s reagent modifies primary amines, located at the N-terminus and on lysine side chains, introducing a sulfhydryl group that is fully compatible with this kit.
These kits utilize Tube-O-Reactor™, which allows for the reactions to be performed in a single tube, with no loss of essential reagents and minimum hands on time and effort.
- Couple to Amine or Sulfhydryl groups
- Simple to use, single step reaction
- Suitable for all carrier proteins
- Uses Tube-O-Reactor™ for minimum hands on time
- Coupling of peptides to carrier proteins
- Suitable for antibody production