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) 可特异性标记掺入健康人全血中表达 PSMA 的 LNCaP 前列腺癌细胞，标记效率达 93.8%，且不会与正常血细胞发生非特异性结合，其与 PSMA 的亲和力约为 8 nM^[1]。
DUPA-FITC (100 nM; 1 h) 可特异性结合 PSMA 阳性的人前列腺癌细胞 LNCaP，而过量的 PSMA 抑制剂 PMPA 可阻断该结合作用^[2]。
DUPA-FITC (100 nM; 1 h) 可特异性结合 PSMA 阳性的人前列腺癌细胞 LNCaP 并被其内化，酸性细胞内内体中会发生荧光淬灭^[2]。

指南 (以下为我们推荐的实验方案。本方案仅作为参考指南，具体操作应根据您的实际需求进行调整)。
1. 细胞制备：制备 PSMA 阳性细胞 (如 LNCaP 或 22Rv1)，或通过密度梯度离心从血液样本中分离出富含 CTC 的组分。
2. 洗涤：用结合缓冲液 (如含 1% BSA 或 0.1% 叠氮化钠的 PBS) 洗涤细胞两次，去除可能干扰结合反应的多余血清蛋白。
3. 孵育：在结合缓冲液中，将细胞与 DUPA-FITC 探针共同孵育。常用浓度范围为 10 nM-100 nM，具体研究可根据探针亲和力调整浓度。
孵育时间与温度：37°C 孵育 1-2 小时，使探针充分结合并发生部分内化；若仅进行细胞膜表面染色，可在冰上孵育 1 小时。
4. 洗涤：用预冷的结合缓冲液洗涤细胞 3 次，去除未结合的游离探针。
5. 检测：采用流式细胞术 (FITC 通道) 或荧光显微镜对细胞进行分析检测。

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

1040.10

C₅₁H₅₇N₇O₁₅S

1123684-41-2

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]