Product Description
CellMosaic® has designed this PerKit™ HRP−Oligo conjugation kit to prepare single-labeled HRP−Oligo conjugates with linkers and conjugation chemistry that retains the activity of HRP. The same configuration and chemistry have been tested for HRP-oligos with different oligo sequences at a few different diagnostic companies since 2011.
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This HRP-Oligo Conjugation kit works with protected thiol-modified oligos (disulfide form). The customer supplies the thiol-modified oligos containing 3’ C6 disulfide linker or 5’ C6 disulfide linker, which are readily available from many commercial oligo suppliers. Using the kit components, the customer reduces the disulfide bond of the oligo, followed by reaction of the thiol-oligo with activated horseradish peroxidase (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 is an upgraded and improved replacement kit with slightly different components and workflow as the first-generation kit (Cat# CM53402). This kit has multiple configurations for various lengths of oligos and can be used to conjugate 2 to 5 nano-mole (nmol) of oligo. Table 1 provides the catalog numbers for various kit configurations.
Table 1: Configurations of the PerKit™ HRP−Oligo conjugation kit (CM53408)
|
Configuration |
No. of Reactions |
Catalog No. |
|
1. Labeling for short oligos (5-30 bases) |
1 |
CM53408.1x1 |
|
3 |
CM53408.1x3 |
|
|
2. Labeling for medium oligos (31-59 bases) |
1 |
CM53408.2x1 |
|
3 |
CM53408.2x3 |
Requirements for disulfide-oligo
- Total amount: 2 to 5 nano-mole oligo content as measured by UV.
- HPLC purified with disulfide-oligo content ≥90% by C18 HPLC.
- ≥5- bases (oligos longer than 60 bases may result in lower loading and high impurities).
Key Features of this HRP-Oligo Conjugation Kit
-
High quality maleimide activated HRP for the conjugation: >99% purity and >200 units/mg protein activity.
- A single purification step affords around 85-90% of single labeled HRP-oligo conjugates if the quality of the oligo is at par.
- Preparation can be done within a few hours.
- Stable linkage with optimal spacer to ensure no interference between oligo binding and HRP activation.
- All reagents included from preparation to purification.
- Options to choose tailored services at CellMosaic after conjugation:
- You can choose to send your conjugates to CellMosaic for HPLC analysis of the sample or complete removal of trace oligo impurities and unreacted HRP.
Labeling Chemistry
