Luminex xMAP Antibody Coupling Kit - Manual (Page 9)

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Protein Considerations
This kit includes a specially formulated Activation Buffer (pH 6.0) which is suitable for
most antibodies.
The protein to be coupled must be free of sodium azide, bovine serum albumin (BSA),
glycine, tris(hydroxymethyl)aminomethane (Tris), glycerol, or amine-containing additives
and should be suspended in phosphate buffered saline (PBS), pH 7.4.
A number of buffers can be used successfully in this coupling reaction. Generally, the pH
at which a coupling reaction occurs should be compatible with the solubility of the protein
of interest. This should be considered when coupling different proteins.
Additionally, coupling efficiency will vary depending on a variety of factors, including type
of antibody, quantity of antibody, quantity of microspheres, etc. As such, it is
recommended that several quantities of antibody input be tested to optimize the coupling
reaction and functionality in the final assay. If coupling for the first time, 2-5μg of antibody
per 1x10
6
microspheres is generally a good starting input.
The amount of antibody to use in the coupling reaction will depend on the quantity of
coupled antibody necessary to promote optimal binding of the desired target molecule.
Kit Contents
The following items are included in this kit:
NOTE: The EDC reagent is a Thermo Scientific product and is manufactured for
Luminex for use in this kit. For questions regarding the use of this product with this
kit, please contact Luminex technical support.
Component Part Number Volume/Mass Quantity
EDC Reagent 11-40144
(ThermoSci #77149)
10 mg 1
Sulfo-NHS 11-25169 250 μL 1
Activation Buffer
(green cap)
11-25171 45 mL 1
Wash Buffer 11-25167 30 mL 1
1.5 mL tubes 11-00277 n/a 10
Disposable pipettes 11-00321 n/a 20