Luminex xMAP Antibody Coupling Kit Manuel d'utilisateur télécharger pdf (Page 22)

Page 18 of 20

Low/No

signals

The reagents may not

have been stored

correctly

Store all reagents as recommended.

EDC may not have

been stored correctly

or rehydrated long

before use

EDC must remain in the pouch it is

shipped in until ready to use.

EDC must be used immediately in the

coupling procedure after rehydration.

The stock protein

concentration is

possibly low

Make sure the protein concentration is

correct and the recommended amount

is coupled to the microspheres.

The stock protein

suspension possibly

contains foreign

proteins, azide,

glycine, Tris or some

primary amines

Dialyze the protein of interest to

remove the competing substances.

The reaction volumes

and/or mixing method

may be incorrect

The final reaction volume for the 2 hr

incubation is critical for successful

coupling. Refer to the protocol. End-to-

end mixing during the 2 hr incubation

is very important. Please use an

optical rotator.

The anti-species

detection antibody

may not be correct

Make sure the anti-species detection

antibody is correct, for example if you

are using a mouse monoclonal to

couple to the microspheres, use an

anti-mouse detection antibody.

Make sure the anti-species detection

antibody has the appropriate label on

it, either biotin or phycoerythrin.

Problem Possible Cause(s) Recommendations

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