Product Description
Alkaline phosphatase (AP) is a hydrolase enzyme responsible for removing phosphate groups from many types of molecules, such as nucleotides and proteins. It is used to detect target proteins through chromogenic, chemiluminescent, or fluorescent outputs. CellMosaic® is a preferred choice for diagnostic companies to source highly pure and single-labeled AP-oligo conjugates. This routine synthesis service is designed for the small-scale preparation of purified and characterized AP–oligo conjugates, with one oligo per AP. This single-labeled conjugate is especially suitable for diagnostic applications with reliable performance.
An amine-modified oligo can be supplied by the customer or purchased by CellMosaic if the sequence is provided. CellMosaic delivers the final conjugate with SEC-HPLC analysis data.
Customers can order online by providing oligo information and selecting from various reaction scales, linker types (stable or releasable), and additional optional services.
What's included in the Base Price:
- Labor and materials (excluding antibody and oligo).
- Initial QC and quantitation of the starting oligo by C18 HPLC to check the purity.
- In-process monitoring of the oligo modification and AP-oligo conjugation.
- Cartridge-type purification to remove any unreacted AP.
- Final product analysis by AEX HPLC to determine purity, aggregation profile, concentration, percentage of unreacted antibody, residual oligo.
Delivery: Final product is supplied in Tris buffer in a single tube along with a COA that includes SEC-HPLC data, concentration, and oligo loading.
Timeline: Approximately 2-3 weeks after receiving the starting antibody and oligo.
Selections and Add-Ons:
1. Reaction Scale: Choose between 5 nmol and 10 nmol oligo labeling scale. Typical recovery is 30-70%, depending on the oligo properties. If you do not see the desired scale, please contact us for a custom quote.
2. Oligo Length: Please indicate the number of nucleotides in your oligo so that appropriate handling and conditions can be applied.
3. Optional AEX Analysis: AEX-HPLC provides charge-based analysis of AOCs and often offers better resolution. If conjugate peaks are separable, AEX can be used to calculate overall oligo loading. Three samples are generally required for this analysis (oligo, antibody, and AOC). [Click here for more information].
4. Optional AEX HPLC purification: For project requires complete removal of unreacted oligo and AP, AEX HPLC is highly effective. After purification, conjugate will be reanalyzed by SEC HPLC to determine the concentration.
Requirements for Amine Oligo:
- Type: Standard amine DNA oligo with deoxyribose–phosphate backbone. For modified backbones such as phosphorothioate (PS), locked nucleic acids (LNA), RNA, or morpholino oligos, please contact us for a quote.
- Purity: HPLC-purified and lyophilized amine oligo with ≥90% amino-oligo content by C18-HPLC. No additional components containing primary/secondary amines or thiol groups should be present.
- Amount: 15 or 30 nmol oligo (3 x amount of choosing scale) (Measured by UV.) Unused materials can be returned upon request
- Information: Provide as much detail as possible — amount, molecular weight, sequence, extinction coefficient — and attach QC data from your oligo vendor. Sequence information is required if CellMosaic is to order the oligo.
Example 1: AP-Oligo Prepared at CellMosaic with Cartridge-Type Purification
Oligo: HPLC-purified 23mer oligo with 5’ NH2-C6 modification
Specification of the final conjugate:
% of AP/Oligo (1:1): 93% Unreacted AP: N/A
Unreacted oligo: N/A% Conjugate recovery: 52% recovery
Figure 1: SEC-HPLC profiles of maleimide activated AP (blue trace), oligo (green trace), and cartridge-type purified product (without HPLC purification) (red trace) detected at 220 nm.
Example 2: AP-Oligo Prepared at CellMosaic with HPLC Purification
Result: >99.9% pure by anion exchange HPLC, characteristic UV spectrum of AP (280 nm) and oligo (260 nm).
