You can try peptide covalent immobilization onto Nunc CovaLink NH plates. In this case you can use peptides of any length for ELISA with paying no attention on their ability to adsorb onto ELISA plates.

Another method is the use of biotinylated peptides that can be attached onto Streptavidin- or Neutravidin-coated plates. Please take care of a linker that should be put between your B-epitope and biotin, since the biotin-binding pocket of streptavidin is rather deep. The linker assures the free presentation of your B-epitope and its accessibility to antibodies.

We employed both methods and found both suitable, depending on the peptide terminus that should be accessible to antibodies, presence of free alpha-amino- or -carboxy, or SH-groups in the peptide, availability of biotin-linked peptides, etc. 

You can also prepare a conjugate of your peptide with a carrier protein. If you obtained your antipeptide antibodies via using the peptide-protein conjugate, then you should conjugate your peptide with another carrier protein and by using another conjugation reagent, in order to avoid the detection of anti-carrier and anti-linker (if the ones exist) antibodies. 

I coat at 10 microgram per ml in PBS using the Nunc Maxisorp 96 well plates.

You can try peptide covalent immobilization onto Nunc CovaLink NH plates. In this case you can use peptides of any length for ELISA with paying no attention on their ability to adsorb onto ELISA plates.

Another method is the use of biotinylated peptides that can be attached onto Streptavidin- or Neutravidin-coated plates. Please take care of a linker that should be put between your B-epitope and biotin, since the biotin-binding pocket of streptavidin is rather deep. The linker assures the free presentation of your B-epitope and its accessibility to antibodies.

We employed both methods and found both suitable, depending on the peptide terminus that should be accessible to antibodies, presence of free alpha-amino- or -carboxy, or SH-groups in the peptide, availability of biotin-linked peptides, etc. 

You can also prepare a conjugate of your peptide with a carrier protein. If you obtained your antipeptide antibodies via using the peptide-protein conjugate, then you should conjugate your peptide with another carrier protein and by using another conjugation reagent, in order to avoid the detection of anti-carrier and anti-linker (if the ones exist) antibodies. 

Dear Kazuo Ohnishi,

Use Peptide attached onto Neutravidin coated plates, it will give best results for your condition and it's best recommended method.

You can also try NH-Nunc CovaLink. it also good.

Hope you got.,

Good Luck & Regards

R.Selva...

The answers given above are fine. Apart from that, i tried with takara peptide coating kit. its ok. Alternatively, you can coat peptides directly with carbonate-bicarbonate buffer, pH 9.6 for me. 37 degrees celsius for 2 hrs. check hydrophobicity of peptide and other parameters too for pH adjustment. It should work fine.  Use Nunc maxisorp microplates

Its interesting to see a number of strategies working for a perceived difficult problem. Has anybody knowledge on the reliability of the various techniques?

For example, when working with peptide pools, individual adjustment of pH is rarely feasible and its important to know how large a fraction - on average - this works for?

For peptide pools, biotin labelling of peptides is the most reliable technique, since it does not depend on peptide structures and properties. Another approach is to use peptides synthesized on solid-phase by using non-cleavable linkers. Both approaches were well developed by Mimotopes and PEPSCAN companies. The use of biotinylated peptides has an advantage that peptides can be checked and purified, if necessary (although it is usually not the case for large peptide sets), before being analyzed.