Can someone suggest how to improve stability of protein in ELISA kit when coated in ELISA plate?
It really depends upon what type of protein or antigen you are coating on the micro titration plates. In general after coating with the antigen the plates should be blocked with other proteins like albumin, ovalbumin, gelatine or other protein which not only block the uncoated space to prevent non-specific reaction and high background of the reaction but also helps to stabilize your antigen or may help as a cryostat. In many cases such plates can be kept for about 6 – 12 monthes even at +4OC. for further details please send un e-mail request
It is a vague question. Did you keep the coated plate in -20C, with desicant, during that time? Also, did you put Na azide in it? Na azide can inhibit HRP activity if you use HRP as the reporter. Without any description in your method makes it hard to decipher.
It really depends upon what type of protein or antigen you are coating on the micro titration plates. In general after coating with the antigen the plates should be blocked with other proteins like albumin, ovalbumin, gelatine or other protein which not only block the uncoated space to prevent non-specific reaction and high background of the reaction but also helps to stabilize your antigen or may help as a cryostat. In many cases such plates can be kept for about 6 – 12 monthes even at +4OC. for further details please send un e-mail request
Alberto Medina is right , we need more information as to what sort of plate it is Precoated or do you coat those plates and the coating buffer pH .Advisable storage temperature is 4 degress .
If you are coating the plate yourself, you may seal/wrap the plate with plastic before incubation at 4 degrees.... Yes depending on other details of information as others have already said.
Good luck.
You can use preservative in you coating solution.for example sodium azid or thimorsal .but i think the best method is selling your plate in aluminum foil with vacuum and silica gel.
I use blocking solution(0.05% Tween 20 +BSA 1%)
I use coating buffer( carbonate/bicarbonate buffer at pH >9)
and storage in 4 degree for 16 hours
I use plate(Grainer bio one)
Even closing the plate with a plate lid and storing them in a moist box /container at 4 degrees serves good , we have been doing it and thats the best we have found :)
1. The really best method is to keep the plates frozen after removing as much of the coating buffer. No washing, no detergents.
2. If you would like to store your coated plates at room temperature, you need a blocking step (1% w/v BSA in PBS), washing once with PBS (no detergents) and than the stabilzing buffer PBS + 0.1% ... 1% w/v BSA + 1% Mannitol + 1% Glucose. Incubate this for 1h, remove as much as possible, let it dry (best in a climate cabinet), make sure that all liquid is gone, than you can seal the plates. Each with a small dry package ...
or go directly to 1.
tank you for your answer
Mr.Stephan Kiessig ...?what is the meaning of ...?
Hi Ali, "..." is "-". Take it a a potential range of concentrations.
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