Product Description
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| SDS | USER MANUAL |
CellMosaic® has designed this personalized immobilization kit to work with small molecules containing one carboxylic acid functional group. The user supplies the small molecule acid, then, using the kit components, converts the carboxylic acid to an activated N-hydroxysuccinimide (NHS) ester, followed by reaction with amine-modified agarose beads.
This kit is custom made to order.
Note: Some of small molecules may contain other functional groups that can interfere with this reaction, such as amine. We recommend that customers first email information about their small molecule before placing an order for this kit so we can provide insight into the efficacy of this kit and potentially present alternative options.
Requirement for small molecule:
- Preferably > 90% pure
- Absence of primary or secondary amine groups
- Non-hindered aliphatic carboxylic acid
- For molecule containing aromatic carboxylic acid, hindered aliphatic carboxylic acid, or hydroxyl groups, please consult CellMosaic prior to conducting the experiment
Key Features of this SepSphere™ Immobilization Kit:
- Offers a simple and easy way to label small molecules with carboxylic acid
- Fast and easy preparation: 3 h preparation and less than 30 minutes hands-on time
- Stable linkage (no leakage of the small molecules from the resin under acid or base condition)
- All reagents and supplies include for preparation
- Use of agarose beads with optimized amount of amine functional groups for loading to preserve the nature of the agarose beads
Labeling Chemistry
Specification
- Matrix: agarose 4B (source: GE Life Science)
- Particle size: 45-165 um
Typical Kit Performance Data (LC Analysis, CellMosaic®)
Small molecule information: Fmoc-Gly-OH
Figure 1: C18 HPLC/UV analysis of Fmoc-Gly-OH before (sample C1) and after immobilization (sample C3) (detected at 254 nm). Left inset: UV spectrum of Fmoc-Gly-OH peak 8.63 min. Right insert: expansion of the HPLC profile from 8.0 to 9.6 min.
Calculate Loading: (6313.73-26.7787)x10/6313.73= 9.96 µmole per mL
Figure 2: C18 HPLC/UV analysis of deprotected resin. Inset: UV spectrum of Fmoc-piperidine adduct at 7.662 min.
Experiment for analyzing the Fmoc group loaded onto the resin: After immobilization of Fmoc-Gly-OH, ~100 µL of beads were removed and added into 200 µL of 20% piperidine in NMP. After mixing for 15 minutes, 5 µL were removed for HPLC analysis. The HPLC profile is identical to the profile obtained from Fmoc-Gly-OH deprotection, confirming that Fmoc-Gly-OH is loaded onto the resin.
