Home > Community > Which is the best microplate for anti-peptide ELISA?
Upvote

30

Downvote
+ Elisa
+ Peptide chemistry
+ Immobilization
+ Immunoassay
+ Peptides
Posted by
Mike Oc

Which is the best microplate for anti-peptide ELISA?

Donald Hubbs  Follow

do have somebody experience with AGID using syn. peptides? on standard polysterone PD?

More

Upvote

VOTE

Downvote
Carlos Torres  Follow

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. 

More

Upvote

VOTE

Downvote
Asim Qureshi  Follow

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?

More

Upvote

VOTE

Downvote
Brian Lawrence  Follow

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. 

More

Upvote

VOTE

Downvote
Curt Weinstein  Follow

Conditions for protein, peptide, and virus coating. Work fantastically well:
Article Antibodies against a preselected peptide recognise and neutr...


More

Upvote

VOTE

Downvote
Donna Bianca  Follow

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. 

More

Upvote

VOTE

Downvote
Billie Lusk  Follow

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

More

Upvote

VOTE

Downvote
Chris Huey  Follow

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

More

Upvote

VOTE

Downvote
Alex Voge  Follow

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

More

Upvote

VOTE

Downvote
Blake Senftner  Follow

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...

More

Upvote

VOTE

Downvote