FITC-UEA-I（荆豆凝集素I-FITC标记）

搜索关键词：

FITC-UEA-I 绿色荧光标记荆豆凝集素I；Anti-H blood group specificity 抗血型H特异性；Lectin Binding 凝集素结合；Endothelial cells内皮细胞；DiI-Ac-LDL 红色荧光探针标记乙酰化LDL；EPC（Endothelial Progenitor Cell）内皮祖细胞；

产品订购：

【温馨提示】：见我司整理的内皮祖细胞（EPC）双荧光鉴定产品专题。

FITC-UEA-I基本描述：

荆豆凝集素，英文全名Ulex Europaeus Agglutinin，缩写UEA，特异性抵抗H血型，常见两种凝集素类型：UEA I结合L-岩藻糖，UEA II结合N,Nˊ-二乙酰壳二糖。最初研究发现UEA I分子量为170kDa，后续研究显示UEA I中性或碱性pH下可能会聚合，而正确分子量为68kDa。

本品为FITC荧光标记的UEA I，即FITC-UEA-I（FITC-UEA-1），经凝胶管柱层析法纯化所得，Ex/Em=490/520nm。结合含α-连接岩藻糖的许多糖蛋白和糖脂，比如ABO血型糖偶联物。优先结合血型O细胞，用于确定分泌型状态。还可用做一种优异的人内皮细胞标志物。适用于免疫细胞/免疫荧光（IC/IF）和糖生物研究。

FITC-UEA-I主要应用：

FITC-UEA-I（FITC-UEA-1），最常见的应用就是联合DiI-Ac-LDL（红色荧光标记乙酰化LDL）一起，用来标记定向诱导培养的单个核细胞（MNCs），以鉴定成功分化的内皮祖细胞（Endothelial Progenitor Cells，EPCs），这是目前来说最成功的最普遍的从功能学角度来鉴定EPCs的方法。

EPCs鉴定-双标记方法【更多内容见官网——内皮祖细胞的双标记染色】

步骤：贴壁细胞先用12 μg/ml DiI-Ac-LDL（货号：MP6013-500UG）37°C孵育4h。PBS清洗细胞，细胞清洗（5min/次，3次），然后用2%多聚甲醛固定10min。之后用10 μg/ml FITC-UEA-1（货号：MP6308-500UG）室温孵育1h。共聚焦显微镜观察染色效果。  

图1：荧光显微镜（40×物镜）双荧光标记观察鸡EPCs对探针的吸附能力，结果显示A：相差；B：DiI-Ac-LDL阳性（红色）；C：FITC-UEA-1阳性（绿色）；D：DiI-Ac-LDL阳性+ FITC-UEA-1阳性（双阳性，黄色荧光）；E：合并图片；

文献来源：Chunyu Bai, et al. Characterization of vascular endothelial progenitor cells from chicken bone marrow. BMC Vet Res. 2012; 8: 54.

基本特性：

1. 英文同义名：UEA I Lectin (FITC); Lectin from Ulex europaeus, FITC conjugate; Fluorescein labeled Ulex Europaeus Agglutinin I; Ulex europaeus Lectin;FITC-UEA-I; FITC-UEA-1; UEA-I-FITC;

2. 分子量：~68 kDa

3. 特异性：岩藻糖、阿拉伯糖

4. FITC/蛋白比率：1.0-1.4（A490/A280）

5. 外观：液体

6. 浓度：2mg/ml

7. 应用：糖生物；IF/ICC（1:50-1:200），具体浓度根据实验体系来摸索；

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 — —Written/Edited by V. Shallan【版权归MKBio懋康所有】

上海懋康生物科技有限公司是一家涉足于生命科学和生物技术领域研究的试剂、仪器和实验室消耗品与实验服务工作，主要从事细胞生物学、植物学、分子生物学、免疫学、生物化学、蛋白组学。生物制药与诊断试剂研发生产等领域。 本公司秉承“以人为本，以诚为信、合同守信”的经营理念。坚持"品质保障"的原则为广大客户提供优质产品。

文献引用：

Quan H et al. LncRNA-AK131850 Sponges MiR-93-5p in Newborn and Mature Osteoclasts to Enhance the Secretion of Vascular Endothelial Growth Factor a Promoting Vasculogenesis of Endothelial Progenitor Cells. Cell Physiol Biochem. 2018;46(1):401-417.

After washing, cells were plated on culture dishes at the density of 5×10 6 and cultured with Microvascular Endothelial Cell Growth Medium-2 (EGM-2 MV) BulletKit (Lonza, Walkersville, MD, USA) in a humidified atmosphere of 5% CO2 at 37°C. Two days later, non-adherent cells were removed by washing with PBS twice and the culture medium was replaced every 3 d. After culturing for 1 d, 7 d, 14 d and 28 d, EPCs were identified by optical microscope observation, immunofluorescence for both Dil-Acetylated Low Density Lipoprotein (Dil-Ac- LDL; MaoKang Biotechnology, Shanghai, China) and FITC labeled Ulex Europaeus Agglutinin 1 (FITC-UEA-1; MaoKang Biotechnology, Shanghai, China), and flow cytometry (FACSCalibur Flow Cytometry, BD, Triangle, NC, USA) for PE-Cy7 conjugated anti-mouse CD34 monoclonal antibody.

Wang, W., Zhang, Y., Hui, H. et al. The effect of endothelial progenitor cell transplantation on neointimal hyperplasia and reendothelialisation after balloon catheter injury in rat carotid arteries. Stem Cell Res Ther 12, 99 (2021). https://doi.org/10.1186/s13287-021-02135-w

During the 8 days of culture, attached EPCs were washed with PBS and subsequently stained with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine-labelled acetylated low-density lipoprotein (Dil-ac-LDL, Maokang Biotechnogy Co., Ltd., Shanghai, China, MP6013) at a concentration of 10 mg/L. After incubation at 37 °C and 5% CO2 for 4 h, cells were fixed with 2% paraformaldehyde (PFA) in PBS for 10 min. Fluorescein isothiocyanate-labelled Ulex europaeus agglutinin (FITC-UEA-1, Maokang Biotechnogy Co., Ltd., Shanghai, China, MP6308) at a concentration of 10 mg/L was added. The cells were examined under a confocal fluorescent microscope (TCS SP5 II, Leica Microsystems, Germany).