CellMosaic has designed this HRP-Oligo conjugate with linkers and conjugation chemistry that retain the activity of HRP. The same configuration and chemistry have been tested for HRP-Oligos with different oligo sequences in several diagnostic companies since 2011.
This HRP-Oligo Conjugation kit works with amine-modified oligos. The customer supplies the amine-modified oligos, which are readily available from many commercial oligo suppliers. Using the kit components, the customer converts the amine-oligo to a thiol-oligo, followed by reaction of the thiol-oligo with activated HRP to generate the HRP-oligo conjugates. The Q-column purification step typically provides the resulting HRP-oligo at greater than 85% purity.
This kit provides materials to label one (CM53401x1) to three oligo samples (CM53401x3). Scale of the reaction: 5 nmol oligo.
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Key Features of this HRP-Oligo Conjugation Kit:
- High quality maleimide-activated HRP for the conjugation: >99% purity and >200 units/mg protein activity
- Optimal maleimide groups per HRP for single label of oligo: 1.2 for typical batch
- A single purification step affords 85~90% of single labeled HRP-oligo conjugates
- Fast preparation: less than 2 h hands-on time
- Easy purification: less than 30 minutes
- Stable linkage with optimal PEG spacer to make sure there is no interference between oligo binding and HRP activation
- No activity loss of the HRP after the conjugation and purification
- All reagents included, from preparation to purification
Typical Kit Performance Data (LC Analysis, CellMosaic)
Oligo information: HPLC-purified 18mer oligo with 5’ NH2-C6 modification
Figure 1: Size exclusion HPLC analysis of the HRP and oligo reaction mixture (before purification).
Figure 2: Size exclusion HPLC analysis of first elution (E1) from Column Q (after purification).