Fluorescent Brightener 28  荧光增白剂28

产品标签

Fluorescent Brightener 28 荧光增白剂28；Calcofluor White M2R；Fungi 真菌；Chitin 几丁质；CAS NO. 4404-43-7；

产品信息

产品描述

荧光增白剂28（Fluorescent Brightener 28），也称为Calcofluor White M2R，常用作真菌染色和细胞活力染色。该染料的用途广泛，包括：1）作为荧光增白剂，工业上用在纤维素和锦纶织物、纸张，和去污剂，肥皂；2）鉴定和研究几丁质的结构和生物合成。

几丁质是自然界中发现第二丰富的多糖，来源于淡水海绵Spongilla lacustris。当与几丁质结合后，荧光增白剂28的荧光得以加强，可用来阐述骨架结构中几丁质的特殊位置；3）用来染色真菌细胞壁；4）用来染色白色念珠菌（Candida albicans）生物膜。

本品以粉末形式提供，CAS NO.4404-43-7，典型工作浓度为0.05 - 0.20%。

作用机制

荧光增白剂28（Fluorescent Brightener 28），同义名Calcofluor White M2R，非特异性结合含β-1,3-连接或β-1,4-连接的多糖，比如几丁质（chitin），β-葡聚糖和纤维素，因此能够用来快速检测细胞壁含此类成分的酵母菌、真菌和寄生生物等。其最大的吸收波长为365nm（UV，紫外激发），最大发射波长为450nm。

产品应用（来自文献）：

1）Gerphagnon M et al. A Double Staining Method Using SYTOX Green and Calcofluor White for Studying Fungal Parasites of Phytoplankton. Appl Environ Microbiol. 2013 Jul;79(13):3943-51. PMID: 23603679 [CFW was used at a 2.5% (vol/vol) final concentration，zoosporic content of sporangia，Fungi]

2）Monheit JG et al. Calcofluor white detection of fungi in cytopathology. Am J Clin Pathol. 1986 Feb;85(2):222-5. [CFW was used at a 0.1% aqueous solution，Fungi in cytology specimens]

3）Domozych D et al. Imaging the Dynamics of Cell Wall Polymer Deposition in the Unicellular Model Plant, Penium margaritaceum. Methods Mol Biol. 2017; 1563:91-105. PMID: 28324604 [Unicellular Model Plant, Penium margaritaceum]

4）Hervé C et al. Monoclonal Antibodies, Carbohydrate-Binding Modules, and the Detection of Polysaccharides in Plant Cell Walls. Methods Mol Biol. 2011; 715:103-13. PMID: 21222079 [0.25% (w/v) Calcofluor White was used, Plant Cell]

5）Demarco D et al. Histochemical Analysis of Plant Secretory Structures.Methods Mol Biol. 2017;1560:313-330. [Calcofluor white solution: dissolve 0.01 g calcofluor white MR2 in 100 mL of distilled water. Store at 4 °C. Plant Cell]

6）Arcones I et al. Monitoring Chitin Deposition During Septum Assembly in Budding Yeast.Methods Mol Biol. 2016;1369:59-72. PMID: 26519305

7）Lee KJ et al. Resin Embedding, Sectioning, and Immunocytochemical Analyses of Plant Cell Walls in Hard Tissues. Methods Mol Biol. 2014; 1080:41-52. PMID: 24132417 [ Calcofluor White: 0.25 % (w/v) in PBS]

8）Green LC et al. Discrimination between Viable and Dead Encephalitozoon cuniculi (Microsporidian) Spores by Dual Staining with Sytox Green and Calcofluor White M2R. J Clin Microbiol. 2000 Oct;38(10):3811-4. PMID: 11015407 [The slides were allowed to dry, quickly fixed in methanol, stained with 5 mg of Calcofluor White M2R/ml of H2O for 5 min, rinsed in H2O, and allowed to dry. Encephalitozoon cuniculi spores]

9）Stagoj MN et al. Microtiter plate assay of yeast cell number using the fluorescent dye Calcofluor White M2R. Biotechniques. 2004 Mar;36(3):380-2. PMID: 15038150 [Calcofluor White M2R stock solution was prepared by dissolving 48 mg solid dye in 10 mL water and filter sterilized. Stock solution can be stored at -20°C for up to 6 months. Calcofluor staining solution in buffer A (25 μM) was made fresh daily and maintained at room temperature in the dark. Cultivate yeast in the microtiter plate.]

产品特性

1）CAS NO：4404-43-7

2）同义名：Calcofluor White M2R卡尔科弗卢尔白M2R；Calcofluor M2R；Tinopal UNPA-GX；

3）分子式：C40H44N12O10S2

4）分子量：916.98g/mol

5）外观：黄色至米黄色粉末

6）Ex/Em：365/435nm

7）溶解性：溶于水（1mg/ml），无水乙醇（2mg/ml），DMSO（50mg/ml），微溶于甲醇

8）化学结构式：

保存与运输方法

保存：室温干燥保存，3年有效。

运输：室温运输。

常见问题（FAQs）

一、如何配制储存液？

Calcofluor White M2R能溶于DMSO（50mg/ml），也能溶于水（1mg/ml），但溶于水可能需借助50% NaOH或KOH来促进完全溶解。

参考方法：称取100mg粉末加入90ml去离子水（蒸馏水），边搅拌边逐滴加入20%（w/v） KOH，直至溶液变得透明。然后用水定容到100ml即得到1mg/ml储存液，0.22um滤膜过滤除菌（可选），按照单次用量分装，-20℃分装冻存。工作液浓度是储存液的10%（v/v）。具体的储存液配制浓度和溶剂请根据实际实验需求来调整。

注意事项

为了您的安全和健康，请穿实验服并戴一次性手套操作。

相关产品

 — —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：

[1]Yu C et al. Nanoprobe-based force spectroscopy as a versatile platform for probing the mechanical adhesion of bacteria. Nanoscale. 2019 Apr 23;11(16):7648-7655. PMID: 30720812

Fluorescent Brightener 28 wasobtained from Maokang Biotechnology (Shanghai, China). For fluorescence staining of extracellularpolymeric substances (EPSs), after the biofilm formation, thePDMS surfaces were incubated with 6 µM SYTO 9 at roomtemperature in the dark for 12 min, followed by a second stain-ing of 0.15 mM fluorescent Brightener 28 for 3 min.

[2]Shengnan Tao et al. Ultra-stable Pickering emulsion stabilized by a natural particle bilayer. Chem. Commun., 2020,56, 14011-14014 https://doi.org/10.1039/D0CC05690A

Calcofluor White was bought from Maokang Biotechnology (China) Co., Ltd. Calcofluor White (SNCs) and Nile Blue A (ZNPs) were used to stain Pickering emulsions. Lasers of 405 nm and 633 nm were used to excite the dyes of Calcofluor White and Nile Blue A, respectively.

[3]Qian Li et al. 2‐Hydroxy‐4‐methoxybenzaldehyde inhibits the growth of Aspergillus flavus via damaging cell wall, cell membrane, manipulating respiration thus creating a promising antifungal effect on corn kernels. https://doi.org/10.1111/ijfs.14617

Calcofluor-White Stain (CAS: 4404-43-7) was purchased from Maokang Biotechnology Co. (Shang-hai, China).

[4]Ya Fei Geng, et al. An innovative role for luteolin as a natural quorum sensing inhibitor in Pseudomonas aeruginosa,Life Sciences,Volume 274,2021,119325,ISSN 0024-3205,https://doi.org/10.1016/j.lfs.2021.119325.

Afterwards, 50 μg/mL of Calcofluor white M2R (Maokang, Shanghai, China), a blue fluorescent marker, was used to visualize the exopolysaccharides in the biofilm matrix by staining for 2 h at room temperature in the dark.

[5]Li, Q., Zhu, X., Xie, Y. et al. o-Vanillin, a promising antifungal agent, inhibits Aspergillus flavus by disrupting the integrity of cell walls and cell membranes. Appl Microbiol Biotechnol 105, 5147–5158 (2021). https://doi.org/10.1007/s00253-021-11371-2

 Calcofluor white (CAS: 4404-43-7) was purchased from Maokang Biotechnology Co. (Shanghai, China) 

[6] JiashunShi et al. Constructing zwitterionic nanofiber film for anti-adhesion of marine corrosive microorganisms. Journal of Materials Science & Technology. Volume 70, 20 April 2021, Pages 145-155

Calcofluor stain was purchased from Maokang Biotechnology Co., Ltd (Shanghai, China). 

[7]QianLi et al. Antifungal efficacy of paeonol on Aspergillus flavus and its mode of action on cell walls and cell membranes. LWT Volume 149, September 2021, 111985

Calcofluor white (CW) (CAS: 4404-43-7) were purchased from the Maokang Biotechnology Co., Ltd 

Fig. 1. Effect of paeonol on cell walls' integrity of A. flavus. (A) SEM observation of the mycelial morphology. Quantification of the content of (B) β-1,3-glucan and (C) chitin in mycelia cell walls. a-c significant difference (P < 0.05) according to Duncan's multiple range test. (D) Distribution of septa in mycelia treated with paeonol after staining with calcofluor white under a fluorescence microscope. Scale bar: 10 μm.

https://ifst.onlinelibrary.wiley.com/doi/abs/10.1111/ijfs.14617

[8]Jing Wang et al. International Journal of Biological Macromolecules Structural and emulsion-stabilizing properties of the alkali-extracted arabinoxylans from corn and wheat brans. International Journal of Biological Macromolecules. Volume 251, 1 November 2023, 126190

Calcofluor white M2R was purchased from Maokang Bio-Technology Company (Shanghai, China).

[9] Wang J, Hou X, Hannachi K, Fan M, Li Y, Qian H, Wang L. A multi-scale approach to arabinoxylan-based emulsions: From molecular features, interfacial properties to emulsion behaviors. Int J Biol Macromol. 2024 Feb;258(Pt 1):128881. doi: 10.1016/j.ijbiomac.2023.128881. Epub 2023 Dec 21. PMID: 38134997.

 Nile red was obtained from Sigma-Aldrich Company (St. Louis, MO, America) and Calcofluor white M2R (CW M2R) was purchased from Maokang Bio-Technology Company (Shanghai, China). 

[10] Wenxiu Teng et al. Modulating the textural and structural characteristics of curdlan-potato protein gel with different concentrations. Food Hydrocolloids. Volume 149, April 2024, 109638

Rhodamine B and calcofluor white were purchased from Sigma-Aldrich, Inc. (Shanghai, China) and MaoKang Biotechnology Co., Ltd. (Shanghai, China).

[11] Du, G., Shao, Y., Luo, B. et al. Compliant Iontronic Triboelectric Gels with Phase-Locked Structure Enabled by Competitive Hydrogen Bonding. Nano-Micro Lett. 16, 170 (2024). https://doi.org/10.1007/s40820-024-01387-4

Calcofluor White Stain was purchased from Shanghai Maokang Biotechnology Co., LTD.

[12] Zhang W, He D, Wei Y, Shang S, Li D and Wang L (2022) Suppression of Aspergillus fumigatus Germination by Neutrophils Is Enhanced by Endothelial-Derived CSF3 Production. Front. Microbiol. 13:837776. doi: 10.3389/fmicb.2022.837776

Methenamine silver staining (MST, BA4094, BASO diagnostics, Inc., China) and Calcofluor white staining (CWS, 4404-43-7, Shanghai Maokang Biotechnology Co., Ltd., China) were used to label A. fumigatus.

[13] Wang Y, Fan J, Zhou Z, Goldman GH, Lu L, Zhang Y. Histone acetyltransferase Sas3 contributes to fungal development, cell wall integrity, and virulence in Aspergillus fumigatus. Appl Environ Microbiol. 2024 Apr 17;90(4):e0188523. doi: 10.1128/aem.01885-23. Epub 2024 Mar 7. PMID: 38451077; PMCID: PMC11022558.

To visualize the chitin content in the cell wall, samples were stained with 5-mg/mL CFW (Maokang Biotechnology, MS4040) in PBS for 1 min. Fluorescence microscopy was conducted after the incubation using a Zeiss Axio imager A1 microscope (Carl Zeiss, Germany) to capture the images.

[14] Qian Li, et al. Antifungal properties and mechanisms of three volatile aldehydes (octanal, nonanal and decanal) on Aspergillus flavus. Grain & Oil Science and Technology. Volume 4, Issue 3, September 2021, Pages 131-140

To investigate the influence of the three volatile aldehydes on the cell wall, Calcofluor White (CW, CAS: 4404-43-7, Maokang Biotechnology Co., Ltd., Shanghai, China) was used for assessment of any changes to cell wall integrity 

[15] Jianming Cheng et al. Application of ultrasound-assisted NaClO in membrane bioreactors membrane cleaning. Desalination and Water Treatment. Volume 264, July 2022, Pages 12-23

Concanavalin A (ConA) and Calcofluor white (CW) were purchased from Maokangbio (Shanghai, China).

[16]Liang Z, Luo Z, Zhang W, Yu K, Wang H, Geng B, Yang Q, Ni Z, Zeng C, Zheng Y, Li C, Yang S, Ma Y, Dai J. Synthetic refactor of essential genes decodes functionally constrained sequences in yeast genome. iScience. 2022 Aug 18;25(9):104982. doi: 10.1016/j.isci.2022.104982. PMID: 36093046; PMCID: PMC9460170.

Calcofluor white Cat# MM1011

[17]Dong Z, An L, Lu M, Tang M, Chen H, Huang X, Hou Y, Shen G, Zhang X, Zhang Y, Xia Q and Zhao P (2021) SPINK7 Recognizes Fungi and Initiates Hemocyte-Mediated Immune Defense Against Fungal Infections. Front. Immunol. 12:735497. doi: 10.3389/fimmu.2021.735497

 About 10 μL protein sample (5 mg mL^-1) and 10 μL heat-killed spores (1×10⁵ conidia mL^-1) labeled with Calcofluor White M2R (MKbio, China) were added to the hemocyte monolayer and incubated at 25°C for 2 h