DUPA-FITC 

DUPA-FITC is a fluorescent reagent targeting PSMA, which specifically binds to prostate cancer cells expressing PSMA without non-specific binding to normal blood cells. DUPA-FITC can label PSMA-expressing prostate cancer cells in whole blood, followed by internalization and trafficking to acidic intracellular endosomes, during which the fluorescence is quenched. When combined with flow cytometry and density gradient centrifugation enrichment, DUPA-FITC enables quantitative analysis of circulating tumor cells in peripheral blood samples from prostate cancer patients^[1][2].

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.

1040.10

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1123684-41-2

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Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. He W, et al. Quantitation of circulating tumor cells in blood samples from ovarian and prostate cancer patients using tumor-specific fluorescent ligands. Int J Cancer. 2008;123(8):1968-1973.  [Content Brief]

  [2]. Kularatne SA, et al. Prostate-specific membrane antigen targeted imaging and therapy of prostate cancer using a PSMA inhibitor as a homing ligand. Mol Pharm. 2009;6(3):780-789.  [Content Brief]