DUPA-FITC

DUPA-FITC (1 μM) specifically labels PSMA-expressing LNCaP prostate cancer cells spiked into healthy human whole blood with 93.8% efficiency and exhibits no nonspecific binding to normal blood cells, with an ~8 nM affinity for PSMA^[1].
DUPA-FITC (100 nM; 1 h) binds specifically to PSMA-positive LNCaP human prostate cancer cells, with binding abrogated by excess PSMA inhibitor PMPA^[2].
DUPA-FITC (100 nM; 1 h) binds specifically to and is internalized by PSMA-positive LNCaP human prostate cancer cells, with fluorescence quenched in acidic intracellular endosomes^[2].

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
1. Cell preparation: Prepare PSMA-positive cells (such as LNCaP or 22Rv1), or isolate the component rich in CTCs from blood samples through density gradient centrifugation.
2. Washing: Wash the cells twice with a buffer solution (such as PBS containing 1% BSA or 0.1% sodium azide) to remove any excess serum proteins that may interfere with the binding reaction.
3. Incubation: The cells are incubated together with the DUPA-FITC probe in the binding buffer. The commonly used concentration range is 10 nM - 100 nM. The specific concentration can be adjusted according to the affinity of the probe for the study.
Incubation time and temperature: Incubate at 37°C for 1-2 hours to allow the probe to fully bind and undergo partial internalization; if only cell membrane surface staining is performed, incubate on ice for 1 hour.
4. Washing: Wash the cells three times with pre-cooled binding buffer to remove the unbound free probes.
5. Analysis: The cells were analyzed and detected using flow cytometry (FITC channel) or fluorescence microscopy.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.