Our CPN™ products can be conjugated to a variety of molecules for specific targeting.
CPNs™ are readily conjugated to proteins using EDC (N-ethyl-N’-dimethylaminopropyl-carbodiimide) to link amine groups (-NH2) on the protein to the carboxyl groups on the surface of the CPN™ (-COOH). Attachment of proteins such as antibodies or streptavidin will generate highly selective CPNs™ for the detection of target molecules.
The brightness of the CPNs™ ensures the detection of target molecules is highly sensitive, with single molecules being detected in flow cytometry and immunocyto/histochemistry.
The affinity of targeting molecules varies greatly and an initial titration of the CPN™:targeting molecule ratio will need to be undertaken. Similarly, final usage dilution of the CPN™-targeting molecule conjugate will need to be determined empirically.
Add 50 μl CPN™ (0.1mg/ml) to 870 μl of water
Add 20 μl of HEPES 1M
Add 20 μl PEG 5% w/v
Add 40 μl streptavidin (1mg/ml) and vortex (several ratios of CPN™:targeting molecule should be tested to identify the optimum conditions, e.g. 10-100 μl targeting molecule (1mg/ml))
Add 20 μl freshly prepared EDC solution 5 mg/ml
Shake mixture for 4 hours at room temperature
Add 20 μl BSA (10% w/v) and leave to shake for a further 30 minutes
The CPN™:streptavidin conjugates can the be purified from the reaction components by precipitating them using a magnet
Re-suspend in 50 μl PBS
PEG (5% w/v)
Streptavidin (1 mg/ml)
EDC [N-(3-Dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride] 5 mg/ml (Freshly make EDC solution and use immediately. Discard any unused.)
BSA (10% w/v)
Phosphate buffered saline
CPNs™ can be used in a variety of in vitro imaging applications to maximise the success of your R&D.