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SDS | USER MANUAL |
This is a personalized conjugation kit (PerKit™) to work with any protein with a MW over 20 KDa and for protein labeling of a small molecule containing carboxylic acid (-COOH) functional groups. Using the kit components, the customer converts the carboxylic acid to an activated N-hydroxysuccinimide (NHS) ester, followed by reaction with the surface amino groups of the protein to form a stable amide bond. The final product is desalted to remove any unreacted small molecule acid.
This kit provides materials to label 15-50 nmole of one protein sample (≥20 KDa)
If you have less than 15 nmole of protein or the MW of protein is less than 20 KDa, you can contact us for a custom conjugation.
If you would like CellMosaic to perform the conjugation instead, please add PerKit™ Custom Conjugation (AS1002) along with this kit.
Shipping policy: Ship at RT, choose overnight (ships Mon.-Thur.) or 2-day delivery (ships Mon.-Wed.) at checkout.
Requirement for Small Molecule
Key Features of this PerKit™
After conjugation: you can choose to send your sample to CellMosaic for HPLC analysis and calculation of the loading.
Labeling Chemistry
Schematic diagram of the workflow for preparing protein-small molecule conjugates starting with 50 nmole of protein (volume of reagents varies if the amount of protein is < 50 nmole)
Typical Kit Performance Data
SM acid information: 6-carboxyfluorescein (20 µmol)
Protein information: Bovine Serum Albumin (BSA) (50 nmol)
Kit Lot number: 5506.S51207
Figure 1: Size-exclusion HPLC analysis of the purified fluorescein labeled BSA from Step D6 using various amounts of Fluorescein NHS ester.
Figure 2: UV/Vis spectrum of the purified fluorescein labeled BSA from Step D6 using various amounts of Fluorescein NHS ester.
Table 1: Summary results of the purity and the average DOL of the fluorescein labeled BSA from Step D6 (Note: Fluorescein is a very hydrophobic compound. A lower DOL was obtained.)
Sample | NHS ester solution from Step B7 (µl) | SEC HPLC Rt (min) 220 nm | ΔRt | % of small molecule after purification | Calc. DOL based on UV/Vis |
BSA | N/A | 16.616 | N/A | N/A | N/A |
R1 | 7.5 | 16.538 | 0.078 | 0 | 1 |
R2 | 15 | 16.466 | 0.150 | 0 | 1.8 |
R3 | 30 | 16.337 | 0.279 | 1.3 | 2.7 |
R4 | 60 | 16.218 | 0.398 | 2.2 | 3.3 |