Phen Green SK diacetate
(Synonyms: 苯绿SK二乙酸酯,PGSK diacetate) 目录号 : GC40243Phen Green SK diacetate (PGSK)是一种重金属荧光指示剂,对Fe2+、Cd2+、Co2+、Ni2+和Zn2+等一系列金属离子具有反应性。
Cas No.:234075-45-7
Sample solution is provided at 25 µL, 10mM.
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Related Biological Data
Extracellular vesicles derived from T lymphocytes promote iron accumulation in macrophages via PKM2. (B) Free iron levels in peritoneal macrophages were measured by a Phen Green SK probe.
Cells suspended in HBSS were loaded with 20 μmol/L Phen Green SK (GC40243-500, Glpbio, USA), a fluorescent iron chelator, for 15 min at 37 ℃.
Redox Biology (2022): 102257. PMID: 35149342 IF: 11.799 -
Related Biological Data
DHA@MIL-101 NRs treated Lewis cells exhibited enhanced cytotoxicity boosted by ROS generation and ferroptosis. C, D Lewis cells were incubated with the DHA, MIL-101 NRs and DHA@MIL-101 NRs for 12 h and intracellular iron ions were detected using a PGSK probe and flow cytometry.
According to the manufacturer’s instruction, PGSK probe (GC40243, GLPBIO Technology Inc, USA) combined with flow cytometry was applied to detect the concentration of intracellular ferric irons.
J Nanobiotechnol 20.1 (2022): 1-19. PMID: 35568865 IF: 10.4345 -
Related Biological Data
Dexmedetomidine produced antioxidant effect and inhibited HR-induced accumulation of intracellular ferrous iron and lipid peroxidation in H9c2 cells. (A-B) Intracellular ferrous iron was detected by Phen Green SK staining.
In brief, H9c2 cells were treated as indicated before 10 μM Phen Green SK (PGSK, Glpbio, USA) was added and incubated for 10 min.
Biomed Pharmacother 154 (2022): 113572. PMID: 35988428 IF: 7.4194 -
Related Biological Data
Effects of DOX triggers activated HSC ferroptosis in vitro. (E) Free iron levels in LX-2were measured by Phen Green SK probe. LX-2 cells were stained in 10 μM Phen Green SK in PBS for 1 h at 37°C and the samples were imaged immediately.
The cells were stained in 10 μM Phen Green SK (Glpbio, United States) in PBS for 1 h at 37°C.
Front Pharmacol 14 (2023): 1135366. PMID: 37007035 IF: 5.9879
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- Datasheet
Protocol for characterization of copper transport on the plasma membrane of CSB cells [1]: |
10 mM Phen Green SK storage solution was prepared with DMSO, and the storage solution was stored away from light at -20℃ or -80℃ after subpackaging, and used up within 2 weeks. Preparation of 80µM Phen Green SK working solution was prepared by preheating serum-free cell medium or PBS. Please adjust the concentration of the working liquid according to the actual situation. 1.Flow cytometry experiments Intact protoplasts isolated from both 0.1 µM (control) and 100µM CuSO4-grown cells were labeled with the copper-sensitive probe Phen Green SK diacetate (PGSK) to demonstrate copper sequestration. Vacuoles were isolated from PGSK-loaded protoplasts to see if the probe could enter this organelle. Samples were analyzed in flow cytometer equipped with an argon-ion laser emitting a 488 nm beam at 15 mW. Green fluorescence was collected through a 488 nm blocking filter, a 550 nm long-pass dichroic filter and a 525 nm band-pass filter. For each sample, 5,000 protoplasts or vacuoles were analyzed at low flow rate. 2.Copper transport by CSB protoplasts with Phen Green SK diacetate Copper transport across the plasma membrane was determined with PGSK using spectrofluorometer with excitation and emission wavelengths set at 506 and 532 nm, respectively. For transport experiments, protoplasts (106) were incubated with 80µM PGSK for 30 min to equilibrate the dye. The loaded protoplasts (100 µl) were added to the assay cuvette with 9 vols. of the mannitol-containing buffer, pH 5.6. After stabilization of the fluorescence signal, different amounts of CuCl2 were added (0.1-3.5 mM, final concentrations). To discriminate the unspecific fluorescence quenching and the quenching derived from copper uptake by protoplasts, the chelators EDTA and BCS were added to the assay medium. The effect of Cu2+ on the hydrolyzed external probe was tested after treatment of PGSK-loaded protoplasts with 0.5% Triton X-100. To study which form of copper ion was taken up by protoplasts, ascorbic acid was added to the assay medium, reducing Cu2+ to Cu+. To assess the effect of various cations on Cu2+ uptake, protoplasts were incubated for 5 min with the chloride forms of Ca2+ (10 mM), K+ (10 mM), Mn2+ (5 mM), Na+ (10 mM) and Zn2+ (10 mM) before the addition of CuCl2. To test how Cu2+ uptake is regulated by copper availability in the medium, cells grown for 6 d were incubated for 8 h with 100 µM CuSO4 or with 20 µM of the chelator BCS. |
Protocol for measure intracellular ferrous iron levels [2]: 1.Cells were treated as indicated before 10 μM Phen Green SK diacetate was added and incubated for 10 min. 2.The cells were washed twice with PBS to remove the excess PGSK. 3.The cells were trypsinized and resuspended in PBS plus 5% FBS. 4.A flow cytometer was used for detection of ferrous iron. Each group was triplicated.
This protocol only provides a guideline, and should be modified according to your specific needs. |
References: [1]. Martins V, Hanana M, et,al. Copper transport and compartmentation in grape cells. Plant Cell Physiol. 2012 Nov;53(11):1866-80. doi: 10.1093/pcp/pcs125. Epub 2012 Sep 5. PMID: 22952251. [2]. Wang Z, Yao M, et,al. Dexmedetomidine attenuates myocardial ischemia/reperfusion-induced ferroptosis via AMPK/GSK-3β/Nrf2 axis. Biomed Pharmacother. 2022 Oct;154:113572. doi: 10.1016/j.biopha.2022.113572. Epub 2022 Aug 18. PMID: 35988428. |
Phen Green SK diacetate (PGSK) represents a fluorescent indicator for heavy metals, demonstrating reactivity towards a range of metal ions encompassing Fe2+, Cd2+, Co2+, Ni2+, and Zn2+. PGSK is more sensitive to quenching by the ferrous ion (Fe2+) than by the ferric ion (Fe3+)[1].The excitation/emission peaks of PGSK diacetate are recorded at 507/532 nm, correspondingly, with its fluorescence being suppressed upon interaction with metal ions. Phen Green SK is a membrane-impermeable fluorophore, sensitive only to free (chelatable) iron, which can be loaded into intact cells[2-4]. This compound has found applications in quantifying iron levels in isolated rat hepatocytes, elucidating the metal ion preferences of divalent metal-ion transporter 1 (DMT1), and tracking copper transportation across P. sativum chloroplast membranes and cells[5-7].
References:
[1]. Shingles R, North M, et,al. Direct measurement of ferrous ion transport across membranes using a sensitive fluorometric assay. Anal Biochem. 2001 Sep 1;296(1):106-13. doi: 10.1006/abio.2001.5209. PMID: 11520038.
[2].Illing AC, Shawki A, et,al.Substrate profile and metal-ion selectivity of human divalent metal-ion transporter-1. J Biol Chem. 2012 Aug 31;287(36):30485-96. doi: 10.1074/jbc.M112.364208. Epub 2012 Jun 26. PMID: 22736759; PMCID: PMC3436370.
[3]. Petrat F, de Groot H, et,al. Determination of the chelatable iron pool of single intact cells by laser scanning microscopy. Arch Biochem Biophys. 2000 Apr 1;376(1):74-81. doi: 10.1006/abbi.2000.1711. PMID: 10729192.
[4]. Rauen U, Petrat F, et,al.Hypothermia injury/cold-induced apoptosis--evidence of an increase in chelatable iron causing oxidative injury in spite of low O2-/H2O2 formation. FASEB J. 2000 Oct;14(13):1953-64. doi: 10.1096/fj.00-0071com. PMID: 11023979.
[5]. Petrat F, Rauen U, et,al.Determination of the chelatable iron pool of isolated rat hepatocytes by digital fluorescence microscopy using the fluorescent probe, phen green SK. Hepatology. 1999 Apr;29(4):1171-9. doi: 10.1002/hep.510290435. PMID: 10094962.
[6]. Shingles R, Wimmers LE, et,al.Copper transport across pea thylakoid membranes. Plant Physiol. 2004 May;135(1):145-51. doi: 10.1104/pp.103.037895. Epub 2004 Apr 30. PMID: 15122011; PMCID: PMC429342.
[7]. Martins V, Hanana M, et,al. Copper transport and compartmentation in grape cells. Plant Cell Physiol. 2012 Nov;53(11):1866-80. doi: 10.1093/pcp/pcs125. Epub 2012 Sep 5. PMID: 22952251.
Phen Green SK diacetate (PGSK)是一种重金属荧光指示剂,对Fe2+、Cd2+、Co2+、Ni2+和Zn2+等一系列金属离子具有反应性。PGSK对亚铁离子(Fe2+)比三铁离子(Fe3+)更敏感[1]。Phen green SK diacetate激发/发射最大值分别为507/532 nm。它作为一种膜不渗透荧光团,仅对游离(可螯合)铁敏感,可被导入到完整细胞中[2-4]。该化合物已被用于定量分离大鼠肝细胞中的铁水平,阐明二价金属离子转运体1 (DMT1)的金属离子偏好,以及追踪铜在豌豆叶绿体膜和细胞中的运输[5-7]。
Cas No. | 234075-45-7 | SDF | |
别名 | 苯绿SK二乙酸酯,PGSK diacetate | ||
化学名 | 3',6'-bis(acetyloxy)-2',7'-dichloro-3-oxo-N-1,10-phenanthrolin-5-yl-spiro[isobenzofuran-1(3H),9'-[9H]xanthene]-5-carboxamide | ||
Canonical SMILES | CC(OC1=CC2=C(C=C1Cl)C3(C(C=CC(C(NC4=CC(C=CC=N5)=C5C6=C4C=CC=N6)=O)=C7)=C7C(O3)=O)C8=CC(Cl)=C(OC(C)=O)C=C8O2)=O | ||
分子式 | C37H21Cl2N3O8 | 分子量 | 706.5 |
溶解度 | DMSO : 50 mg/mL (70.77 mM; Need ultrasonic) | 储存条件 | Store at -20°C |
General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。 储备液的保存方式和期限:-80°C 储存时,请在 6 个月内使用,-20°C 储存时,请在 1 个月内使用。 为了提高溶解度,请将管子加热至37℃,然后在超声波浴中震荡一段时间。 |
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Shipping Condition | 评估样品解决方案:配备蓝冰进行发货。所有其他可用尺寸:配备RT,或根据请求配备蓝冰。 |
制备储备液 | |||
1 mg | 5 mg | 10 mg | |
1 mM | 1.4154 mL | 7.0771 mL | 14.1543 mL |
5 mM | 0.2831 mL | 1.4154 mL | 2.8309 mL |
10 mM | 0.1415 mL | 0.7077 mL | 1.4154 mL |
第一步:请输入基本实验信息(考虑到实验过程中的损耗,建议多配一只动物的药量) | ||||||||||
给药剂量 | mg/kg | 动物平均体重 | g | 每只动物给药体积 | ul | 动物数量 | 只 | |||
第二步:请输入动物体内配方组成(配方适用于不溶于水的药物;不同批次药物配方比例不同,请联系GLPBIO为您提供正确的澄清溶液配方) | ||||||||||
% DMSO % % Tween 80 % saline | ||||||||||
计算重置 |
计算结果:
工作液浓度: mg/ml;
DMSO母液配制方法: mg 药物溶于 μL DMSO溶液(母液浓度 mg/mL,
体内配方配制方法:取 μL DMSO母液,加入 μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入 μL saline,混匀澄清。
1. 首先保证母液是澄清的;
2.
一定要按照顺序依次将溶剂加入,进行下一步操作之前必须保证上一步操作得到的是澄清的溶液,可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Baicalein and luteolin inhibit ischemia/reperfusion-induced ferroptosis in rat cardiomyocytes
Int J Cardiol 2023 Mar 15;375:74-86.PMID:36513286DOI:10.1016/j.ijcard.2022.12.018
Background: Ischemia/reperfusion (I/R) is associated with severe cellular damage and death. Ferroptosis, a new form of regulated cell death caused by the accumulation of iron-mediated lipid peroxidation, has been found in several diseases including I/R injury, which was reported to be suppressed by flavonoids. Baicalein (BAI) and luteolin (Lut) are flavonoids and were shown to reduce the myocardial I/R injury. BAI was found to suppress ferroptosis in cancer cells via reducing reactive oxygen species (ROS) generation. However, the anti-ferroptosis effect of Lut on ferroptosis has not been reported. This study aimed to investigate whether ferroptosis reduction contributes to the BAI- and Lut-protected cardiomyocytes. Methods: This research used erastin, RSL3, and Fe-SP to induce ferroptosis. Cell viability was examined using MTT assay. Annexin V-FITC, CM-H2DCFDA, and Phen Green SK diacetate (PGSK) fluorescent intensity were detected to analyze apoptotsis, ROS levels, and Fe2+ concentrations, respectively. qPCR and Western blot analysis were conducted to detect the levels of mRNA and protein, respectively. Results: Our data show that BAI and Lut protected cardiomyocytes against ferroptosis caused by ferroptosis inducers and I/R. Moreover, both BAI and Lut decreased ROS and malondialdehyde (MDA) generation and the protein levels of ferroptosis markers, and restored Glutathione peroxidase 4 (GPX4) protein levels in cardiomyocytes reduced by ferroptosis inducers. BAI and Lut reduced the I/R-induced myocardium infarction and decreased the levels of Acsl4 and Ptgs2 mRNA. Conclusions: BAI and Lut could protect the cardiomyocytes against the I/R-induced ferroptosis via suppressing accumulation of ROS and MDA.