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以后地位: 首页> 产品中心> 细胞生物学 > 荧光探针与细胞染色 > MKBio FeRhoNox-1 (Fe2+ indicator) 亚铁离子荧光探针
MKBio FeRhoNox-1 (Fe2+ indicator) 亚铁离子荧光探针
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目次号 MX4558-50UG 售价 1300.00元
规格50μg 运输温度 冰袋运输
其他称号RhoNox-1 保管温度 -20℃避光枯燥保管
CAS号N/A 无效期 至多1年
使用亚铁离子探针「偏向定位高尔基体」 订购数目
产品简介:


FeRhoNox-1 (Fe2+indicator) 亚铁离子荧光探针


产品标签

FeRhoNox-1;FerroOrange;Labile ferrous ion 不波动铁(II)离子;Fe2+荧光探针;Ferroptosis 铁殒命;C11 BODIPY 581/591 脂质过氧化探针;


产品信息

产品称号

产品编号                 

规格                 

代价(元)       

FeRhoNox-1 (Fe2+ indicator) 亚铁离子荧光探针  

MX4558-50UG

50μg

1300

FeRhoNox-1 (Fe2+ indicator) 亚铁离子荧光探针

MX4558-100UG

2×50μg

2500

FeRhoNox-1 (Fe2+ indicator) 亚铁离子荧光探针

MX4558-250UG

5×50μg

6150


产品形貌

FeRhoNox-1,也称为RhoNox-1,是一种活泼的荧光探针,特异性检测不波动的铁(II)离子(Fe2+)。一旦与Fe2+反响后,不行逆的天生一种橙色(白色)荧光产品(Absmax=540nm,FLmax=575nm,图1.FeRhoNox-1的光谱特性)。心理浓度下的铁(III)离子(Fe3+)或别的除铁离子以外的二价金属离子都不会使其荧光加强(见图2FeRhoNox-1的选择性).FeRhoNox-1的反响特异性)。FeRhoNox-1具细胞膜浸透性和高选择性,实用于活细胞内Fe2+的检测,偏向定位在高尔基体。

图1. FeRhoNox-1的光谱特性。FeRhoNox-1与Fe2+反响后吸取和发射光谱(上)。FeRhoNox-1在37℃,与Fe2+反响1h后,荧灼烁显加强。最大荧光峰约在575nm。

图2. FeRhoNox-1的选择性。FeRhoNox-1仅与Fe2+反响。


保管与运输办法

保管:-20℃避光枯燥保管,至多1年无效。

运输:冰袋运输。


留意事变

  1. FeRhoNox-1偏向定位在高尔基体,但也大概检测细胞质池内的Fe2+,现在针对这一点未做明白评价。

  2. 荧光染料均存在淬灭题目,请只管即便留意避光,以减缓荧光淬灭。

  3. 为了您的宁静和安康,请穿实行服并戴一次性手套操纵。


利用阐明

1. 必要自行预备的质料

1.1细胞培育级或超纯DMSO(好比:MS4601A-100ML)【激烈发起将高纯的DMSO分装成单次用量保管在极高温冷冻室内,好比-80℃,制止吸潮。降解的DMSO大概会增长FeRhoNox-1的配景信号】

1.2符合的洗濯和察看缓冲液(好比:PBS, pH 7.4;HBSS;等)。不要含酚红。


2. 探针预备

2.1从冰箱取出FeRhoNox-1,置于室温回温至多30min,将其置于微量离心机内低速离心。将瓶内的粉末离心到管底后,再开盖。

2.2往一管FeRhoNox-1(50μg)内参加109μl高质量DMSO,用枪重复吹吸5次或以上,使其完全消融即失掉1mMFeRhoNox-1贮存液。发起单次用完贮存液,若真实用不完,请依据单次用量分装,置于-80℃避光保管。用中性缓冲液来浓缩贮存液。

2.3于正式实行前,用HBSS或其他中性缓冲液来浓缩1mMFeRhoNox-1贮存液到所需事情浓度(好比:5μM),事情液需现配现用,尽快用完。【留意:酸性溶液会氧化FeRhoNox-1,严峻影响探针的服从】。


3. 染色步调

4. 荧光检测

关于荧光引发:通用的绿色引发滤片好比Cy3或四甲基罗丹明(TMR)检测用的滤片。

关于激光引发:532nm或543nm激光器比力合适。发射波长为570nm左右。


相干产品

货号

称号

规格               

MX5211-1MG           

C11 BODIPY 581/591 脂质过氧化荧光探针

1mg

MX4558-50UG

FeRhoNox-1 (Fe2+indicator) 亚铁离子荧光探针

50μg

MX4559-24UG

FerroOrange (Fe2+indicator) 亚铁离子荧光探针

24μg

MX5401-1MG

MitoPerOx Mitochondrial Lipid Peroxidation Indicator线 粒体脂质过氧化探针

1mg

MX5402-1MG

BODIPY 558/568 C12脂质转运荧光探针

1mg


附录F eRhoNox-1的染色示例

I. HepG2细胞

图3. FeRhoNox-1在HepG2活细胞内的成像检测。①-Fe(II):细胞未添加亚铁离子(100μM硫酸亚铁铵);②+Fe(II):细胞加载亚铁离子;③+Fe(II)+Bpy:细胞加载亚铁离子,之后用参加铁离子螯合剂Bpy。图②荧光加强,而图③荧来临低。此后果与FeRhoNox-1特异性检测Fe(II)的特性分歧。

 

 — —Written/Edited by V. Shallan【版权归MKBioag九游会一切】

 

 

上海ag九游会生物科技有限公司是一家涉足于生命迷信和生物技能范畴研讨的试剂、仪器和实行室斲丧品与实行办事事情,次要从事细胞生物学、动物学、分子生物学、免疫学、生归天学、卵白组学。生物制药与诊断试剂研产生产等范畴。 ag九游会承袭“以人为本,以诚为信、条约取信”的谋划理念。对峙"品格保证"的准绳为宽大客户提供优质产品。


援用文献:

[1]

Lu Zhou, Peng Yu, Ting-ting Wang, Yi-wei Du, Yang Chen, Zhen Li, Man-lin He, Lan Feng, Hui-rong Li, Xiao Han, Heng Ma, Hong-bao Liu, "Polydatin Attenuates Cisplatin-Induced Acute Kidney Injury by Inhibiting Ferroptosis", Oxidative Medicine and Cellular Longevity, vol. 2022, Article ID 9947191, 14 pages, 2022.  


 

Method: FeRhoNox-1 fluorescent probe (MX4558) was purchased from Maokang Biotech (Shanghai, China).FeRhoNox-1,which is a turn-on fluorescent probe specific for the detection of labile iron Fe2+, was used to detect intracellular LIP, and the cellular distribution of FeRhoNox-1 was consistent with Golgi [41]. HK-2 cells were grown to confluence in 35 mm laser confocal petri dishes in DMEM, and PD (40 μM) or Fer-1 (1 μM) was added in the absence or presence of cisplatin (20 μM). Cells were incubated with 5 μM FeRhoNox-1 for 1 h prior to assays. Cells were washed twice with PBS before staining nuclei with Hoechst 33342. The fluorescence was immediately observed with a confocal laser-scanning microscope (CLSM, ECLIPSE Ti, Nikon, Tokyo, Japan).


 

[2]

Jin R, Yang R, Cui C, Zhang H, Cai J, Geng B, Chen Z. Ferroptosis due to Cystathionine γ Lyase/Hydrogen Sulfide Downregulation Under High Hydrostatic Pressure Exacerbates VSMC Dysfunction. Front Cell Dev Biol. 2022 Feb 3;10:829316. doi:


10.3389/fcell.2022.829316. PMID: 35186934; PMCID: PMC8850391. 

Method: For FeRhoNox-1 staining, 5 μM of the FeRhoNox-1 staining working solution (MX4558 and MKBIO)was added and incubated in a 37°C, 5% CO2 incubator for 60 min after treatment for 24 h in a hydrostatic pressure chamber. After washing three times with PBS, cells were imaged with a confocal microscope.


 

[3] Zhang X, Ma Y, Ma J, Yang L, Song Q, Wang H, Lv G. Glutathione Peroxidase 4 as a Therapeutic Target for Anti-Colorectal Cancer Drug-Tolerant Persister Cells. Front Oncol. 2022 Jun 3;12:913669. doi: 10.3389/fonc.2022.913669. PMID: 35719967; PMCID: PMC9203854.


Method: FeRhoNox-1 (an Fe2+ indicator, Cat# MX4558) was purchased from MKBio (Shanghai, China). Ferrous Iron Staining

 

Cells were incubated for 1 h at 37°C with FeRhoNox-1 (an Fe2+ indicator) to detect ferrous iron. The cells were then harvested by trypsinization, and the level of ferrous iron was determined by imaging using a confocal microscope or by flow cytometry analysis.

 


[4] Fang J, Yuan Q, Du Z, Fei M, Zhang Q, Yang L, Wang M, Yang W, Yu J, Wu G, Hu J. Ferroptosis in brain microvascular endothelial cells mediates blood-brain barrier disruption after traumatic brain injury. Biochem Biophys Res Commun. 2022 Sep 3;619:34-41. doi: 10.1016/j.bbrc.2022.06.040. Epub 2022 Jun 14. PMID: 35728282.

Method: Intracellular Fe 2+ levels were evaluated by using FeRhoNox-1 probe (MKBio, Shanghai). 24 h after SI or RSL3 incubation, culture medium was washed by PBS and exchanged for HBSS



 Lai Y, Zeng F, Chen Z, et al. Shikonin Could Be Used to Treat Tubal Pregnancy via Enhancing Ferroptosis Sensitivity. Drug Design, Development and Therapy. 2022 ;16:2083-2099. DOI: 10.2147/dddt.s364441. PMID: 35800255; PMCID: PMC9255906.



Method: Labile Iron Pool (LIP) Assay

“Labile iron” (which is primarily in the ferrous (Fe2+) form) is a small, transitional pool of intracellular iron, and commonly termed “LIP”. LIP release was measured using a FeRhoNox-1™ (Fe2+ indicator) fluorescent probe (MKBio, Beijing, China). HTR-8/SVneo cells were plated in six-well plates, loaded with FeRhoNox-1 (5 μM) for 30 min at 37°C and then washed thrice with Hanks’ balanced salt solution. Cells were observed under a fluorescence microscope (Olympus).

 

[5] Cui J, Zhou Q, Yu M, Liu Y, Teng X, Gu X. 4-tert-butylphenol triggers common carp hepatocytes ferroptosis via oxidative stress, iron overload, SLC7A11/GSH/GPX4 axis, and ATF4/HSPA5/GPX4 axis. Ecotoxicol Environ Saf. 2022 Sep 1;242:113944. doi: 10.1016/j.ecoenv.2022.113944. Epub 2022 Aug 1. PMID: 35926411.



[6] 

Method:  Intracellular Fe2+ determination

FeRhoNox-1 (Fe2+ Indicator) fluorescent probe (Maokang Biotechnology Co., Ltd., Shanghai, China) was used to detect intracellular Fe2+ content. The cells inoculated in cell culture dishes were treated separately (Please see 2.9 for details), and then were washed twice with PBS. The FeRhoNox-1 (5 μM) was added in the medium and the cells were incubated for 60 min. Finally, images were obtained under the fluorescence microscope, and Image J version 1.43 u software was used to quantify the fluorescence intensity.


 

{7]Hong H, Lin X, Xu Y, Tong T, Zhang J, He H, Yang L, Lu Y, Zhou Z. Cadmium induces ferroptosis mediated inflammation by activating Gpx4/Ager/p65 axis in pancreatic β-cells. Sci Total Environ. 2022 Nov 25;849:157819. doi: 10.1016/j.scitotenv.2022.157819. Epub 2022 Aug 2. PMID: 35931150.


Method: For cellular Fe 2+ detection, cells were stained with 5 μmol/L FeRhoNox-1 (Fe 2+ indicator, Maokangbio, China) for 60 min at incubator.


 

[8]

Hu Q, Zuo T, Deng L, Chen S, Yu W, Liu S, Liu J, Wang X, Fan X, Dong Z. β-Caryophyllene suppresses ferroptosis induced by cerebral ischemia reperfusion via activation of the NRF2/HO-1 signaling pathway in MCAO/R rats. Phytomedicine. 2022 Jul 20;102:154112. doi: 10.1016/j.phymed.2022.154112. Epub 2022 Apr 22. PMID: 35550220.


Method:The iron level in astrocytes was detected through FeRhoNox-1 (Fe 2+ indicator) (MX4588, Maokangbio, China)

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