WSP-5 (H2S Probe)  硫化氢荧光探针

产品标签

WSP-1 (Washington State Probe-1)；WSP-5；硫化氢（H2S）特异性探针；PAG (DL-propargylglicine) 硫化氢合成抑制剂；Hypotaurine  H2S清除剂；CAS NO：1593024-78-2；

产品信息

产品描述

WSP-5 (Washington State Probe-5)是由M. Xian团队开发的一种含活性二硫化物的荧光探针，特别设计用来快速检测生物样本和细胞内的硫化氢（H2S）。通过基于反应的荧光开启策略，WSP-5选择性且快速与H2S反应，产生一种荧光化合物（Ex=502nm，Em=525nm）。与WSP-1（货号：MX5301-1MG）相比，WSP-5靶向H2S表现出更快的荧光开启速率（60 vs. 130-fold）和更灵敏的检测极限（47 vs. 60 nM）。

参考文献：Peng B et al. et al. Fluorescent probes based on nucleophilic substitution-cyclization for hydrogen sulfide detection and bioimaging. Chemistry. 2014 Jan 20;20(4):1010-6.

产品特性

1）   CAS NO：1593024-78-2

2）   化学名：3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl bis(2-(pyridin-2-yldisulfanyl)benzoate)

3）   同义名：Washington State Probe-5；硫化氢荧光探针；H2S荧光探针；

4）   分子式：C44H26N2O7S4

5）   分子量：822.9

6）   纯度：≥95%

7）   Ex/Em：502/525nm

8）   外观：浅黄色固体

9）   溶解性：溶于DMSO（~25mg/ml）、DMF（~30mg/ml）

10）化学结构式：

保存与运输方法

保存：-20ºC避光干燥保存，至少2年有效。

运输：冰袋运输。

使用方法【见附录使用延伸资料阅读】

1、  储存液制备

将低温保存的产品从冰箱取出，置于室温回温至少20min。低速离心3-5min。之后往瓶内加入适量无水DMSO充分溶解配成15mM储存液（比如，1mg WSP-5（Mw: 591.7）加81μl DMSO）或更低浓度的储存液，请根据实际情况来调整。按照单次用量分装冻存，避免反复冻融，至少3个月稳定。

2、  工作浓度

正式实验前，取15mM或其他浓度储存液用适当缓冲液或培养基稀释到所需工作浓度即可。具体的工作浓度请参考文献或根据自身实验需求来进行优化调整。参考下方一些实验数据，来自公开文献，仅作参考。

应用示例1：HeLa细胞用DMEM+10% FBS培养2天。成像前一天，细胞转移到24孔板内。检测前，贴壁细胞用FBS-free DMEM清洗一遍。对于胞内H2S成像，细胞用含WSP-5（50μM）的FBS-free DMEM于37℃，孵育30min。用PBS（pH 7.4）清洗取出多余探针，然后细胞用溶于PBS buffer（pH 7.4，100μM CTAB）的NaHS（100μM）孵育30min。用PBS（pH 7.4）清洗细胞3遍后，荧光显微镜成像（490nm激发，绿色通道）。

文献来源：Peng B et al. Nucleophilic substitution-cyclization based fluorescent probes for hydrogen sulfide detection and bioimaging. Chemistry. 2014 Jan 20;20(4):1010-6.

应用示例2：SHSY5Y和U87MG细胞用待研究的缓释天然硫化氢供体HBITC（40, 60, 80 μM）或等体积含DMSO的培养基孵育。之后细胞用硫化氢探针WSP-5（50 μM）和CTAB（100μM in PBS）避光孵育20min。用PBS清洗后，荧光显微镜成像检测。文献来源：Jurkowska H et al. New aspects of antiproliferative activity of 4-hydroxybenzyl isothiocyanate, a natural H2S-donor. Amino Acids. 2018 Jun;50(6):699-709.

注意事项

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引用文献

[1]He T et al. Tumor pH-responsive metastable-phase manganese sulfide nanotheranostics for traceable hydrogen sulfide gas therapy primed chemodynamic therapy. Theranostics. 2020; 10(6): 2453–2462.

H2S Fluorescent Probe WSP-5 was purchased from Shanghai Maokang Biotechnology

Co., Ltd. (Shanghai, China). H2S gas released in 4T1 cells was observed by adding a Washington State Probe-5 (WSP-5) H2S fluorescent probe (Figure Figure33H). It is based on 2-pyridyl disulfide fluorescent, which release the fluorophores and turn on the fluorescence by tandem nucleophilic subsitution-cyclization reaction. WSP-5 probe can selectively and rapidly react with H2S in cells to generate a green fluorescence signal

[2]

Longna Li, Yuhao Liu, Shu Wang, Jianxin Zou, Wenjiang Ding, Wenbiao Shen. Magnesium Hydride-Mediated Sustainable Hydrogen Supply Prolongs the Vase Life of Cut Carnation Flowers via Hydrogen Sulfide. Front. Plant Sci., 09 December 2020 | https://doi.org/10.3389/fpls.2020.595376

H2S fluorescent probe 3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-3’,6’-diyl bis(2-(pyridin-2-yldisulfanyl)benzoate) (WSP-5; MKBio, Shanghai, China) was used to monitored endogenous H2S in cut flowers (Peng et al., 2014). The concentrations of these chemicals were selected based on the results of pilot experiments.

[3] Sun Q, Zhong H, Yue Y, Xiong F, Chen L, Peng X, Chen P, Wan C, Yao Z, Zeng Y. Endogenous hydrogen sulfide promotes human preimplantation embryonic development by regulating metabolism-related gene expression. Nitric Oxide. 2022 Mar 1;120:9-15. doi: 10.1016/j.niox.2021.12.008. Epub 2022 Jan 5. PMID: 34995767.

The H 2 S fluorescent probe 3-oxo-3H-spiro [isobenzofuran-1,9′-xanthene]-3′,6′-diyl bis(2-(pyridin-2-yldisulfanyl) benzoate) (WSP-5; Shanghai Maokang Biotechnology Co, Ltd

[4] Jiale Chen et al. H2S-releasing versatile hydrogel dressing with potent antimicrobial, anti-inflammatory, epithelialization and angiogenic capabilities for diabetic wound healing. Chemical Engineering Journal ( IF 13.3 ) Pub Date : 2023-06-07 , DOI: 10.1016/j.cej.2023.143985

 A cell counting kit-8 (CCK-8), Living/Dead cell double staining kit and WSP-5 were supplied by Maokang Biotechnology Co., Ltd. (Shanghai, China). 

[5] Ou Q, Qiao X, Li Z, Niu L, Lei F, Cheng R, Xie T, Yang N, Liu Y, Fu L, Yang J, Mao X, Kou X, Chen C, Shi S. Apoptosis releases hydrogen sulfide to inhibit Th17 cell differentiation. Cell Metab. 2024 Jan 2;36(1):78-89.e5. doi: 10.1016/j.cmet.2023.11.012. Epub 2023 Dec 18. PMID: 38113886.

H₂S probe staining

Endogenous H₂S production of cells and apoVs was detected by Washington State Probe-5 (WSP-5, MKBio, MX5302), which is based on the dual-nucleophilicity of H₂S and selectively reacted with H₂S to generate a green fluorescence signal.⁵⁴ Cells and apoVs were cultured with WSP-5 for 30 mins and washed with PBS three times. The fluorescent of WSP-5 was detected by confocal microscopy.

 — —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。