Introduction – Luminex xMAP Antibody Coupling Kit User Manual
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Introduction
The xMAP Antibody Coupling (AbC) Kit contains all of the reagents necessary to
covalently couple antibodies to Luminex MagPlex microspheres (beads) in approximately
three hours. This kit can also be used to couple other proteins to Luminex microspheres
but due to the large diversity in protein composition, coupling performance with other
proteins is not guaranteed. For more information on protein coupling please visit:
Coupling is achieved through carbodiimide reactions involving the primary amino groups
on the antibody, or protein of choice, and the carboxyl functional groups on the
microsphere surface.
This kit is configured for a one-time use
*
and contains enough reagent to couple as few as
2.5x10
6
or as many as 50x10
6
microspheres. The kit can be used to perform as many as
ten individual coupling reactions at scales of 2.5x10
6
to 5x10
6
microspheres per
reaction
**
; or as many as four reactions at scales of up to 12.5x10
6
microspheres per
reaction.
IMPORTANT:
* Luminex strongly recommends that the EDC reagent be discarded after one use.
** Each coupling reaction requires a minimum of 2.5x10
6
microspheres.
The coupled microspheres can then be used with a Luminex xMAP instrument to develop
monoplex or multiplex assays.
This kit is ideal for use in two modes:
•
Coupling optimization and assay optimization: For users developing a new assay,
the kit will allow them to perform up to 10 small scale coupling reactions to test
multiple concentrations of antibody with multiple bead regions.
•
Small-to-medium scale assay manufacturing: For users who have already
developed and optimized an assay, the kit will allow them to couple up to 50 million
beads at once to meet their routine usage needs.
Scale
(beads/rxn)
Number of
Reactions
5x10
6
or less
**
10
Up to 12.5x10
6
4