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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Phalloidin-iFluor™ 750 Conjugate/23130/300 Tests
产品编号:23130
市  场 价:¥85560.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$4278.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Phalloidin-iFluor™ 750 Conjugate/23130/300 Tests
商品介绍
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Ex/Em (nm) 752/778
MW ~3300
CAS # N/A
Solvent DMSO
Storage F/D/L
Category Cell Biology
Labeling Cells
Related Biochemical Assays
This NIR fluorescent phalloidin conjugate selectively binds to F-actins with much higher photostability than the fluorescein-phalloidin conjugates. Used at nanomolar concentrations, phalloidin derivatives are convenient probes for labeling, identifying and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. Phalloidin binds to actin filaments much more tightly than to actin monomers, leading to a decrease in the rate constant for the dissociation of actin subunits from filament ends, essentially stabilizing actin filaments through the prevention of filament depolymerization. Moreover, phalloidin is found to inhibit the ATP hydrolysis activity of F-actin. Phalloidin functions differently at various concentrations in cells. When introduced into the cytoplasm at low concentrations, phalloidin recruits the less polymerized forms of cytoplasmic actin as well as filamin into stable "islands" of aggregated actin polymers, yet it does not interfere with stress fibers, i.e. thick bundles of microfilaments. The property of phalloidin is a useful tool for investigating the distribution of F-actin in cells by labeling phalloidin with fluorescent analogs and using them to stain actin filaments for light microscopy. Fluorescent derivatives of phalloidin have turned out to be enormously useful in localizing actin filaments in living or fixed cells as well as for visualizing individual actin filaments in vitro. Fluorescent phalloidin derivatives have been used as an important tool in the study of actin networks at high resolution. AAT Bioquest offers a variety of fluorescent phalloidin derivatives with different colors for multicolor imaging applications.
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This protocol only provides a guideline, and should be modified according to your specific needs.

1. Prepare 1000 X Phalloidin DMSO stock solution: by adding 30 uL of DMSO into the powder form vials.

2. Prepare 1X Phalloidin conjugate working solution: by adding 1 uL of 1000X Phalloidin conjugate DMSO solution to

1 mL of PBS with 1% BSA.

Note 1: The unused 1000X DMSO stock solution of phalloidin conjugate should be aliquoted and stored at -20 oC. protected from light.

Note 2: Different cell types might be stained differently. The concentration of phalloidin conjugate working solution should be prepared accordingly.

3. Stain the cells:

3.1    Perform formaldehyde fixation. Incubate cells with 3.0–4.0 % formaldehyde in PBS at room temperature for 10–30 minutes.

Note: Avoid any methanol containing fixatives since methanol can disrupt actin during the fixation process. The preferred fixative is methanol-free formaldehyde.

3.2    Rinse the fixed cells 2–3 times in PBS.

3.3    Optional: Add 0.1% Triton X-100 in PBS into fixed cells (from Step 2.2) for 3 to 5 minutes to increase permeability. Rinse the cells 2–3 times in PBS.

3.4    Add 100 uL/well (96-well plate) of phalloidin conjugate working solution (from Step 1) into the fixed cells (from Step 2.2 or 2.3), and stain the cells at room temperature for 20 to 90 minutes.

3.5    Rinse cells gently with PBS 2 to 3 times to remove excess phalloidin conjugate before plating, sealing and imaging under microscope.

References & Citations
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Biomaterial Surface Can Modify HUVEC Morphology and Inflammatory Response by Regulating MicroRNA Expression
Authors: Shuangying Gu, Baoxiang Tian, Weicong Chen, Yue Zhou
Journal: Journal of Biosciences and Medicines (2017): 8

Cell-Permeable, MMP-2 Activatable, Nickel Ferrite and His-tagged Fusion Protein Self-Assembled Fluorescent Nanoprobe for Tumor Magnetic Targeting and Imaging
Authors: Lu Sun, Shuping Xie, Jing Qi, Ergang Liu, Di Liu, Quan Liu, Sunhui Chen, Huining He, Victor C Yang
Journal: ACS Applied Materials & Interfaces (2017)

DNA Double-Strand Breaks Induce the Nuclear Actin Filaments Formation in Cumulus-Enclosed Oocytes but Not in Denuded Oocytes
Authors: Ming-Hong Sun, Mo Yang, Feng-Yun Xie, Wei Wang, Lili Zhang, Wei Shen, Shen Yin, Jun-Yu Ma
Journal: PloS one (2017): e0170308

DNA Double-Strand Breaks Induce the Nuclear Actin Filaments Formation in Cumulus-Enclosed Oocytes but Not in Denuded Oocytes
Authors: Ming-Hong Sun, Mo Yang, Feng-Yun Xie, Wei Wang, Lili Zhang, Wei Shen, Shen Yin, Jun-Yu Ma
Journal: PloS one (2017): e0170308

Enhanced bovine serum albumin absorption on the N-hydroxysuccinimide activated graphene oxide and its corresponding cell affinity
Authors: Kun Xiong, Qingbo Fan, Tingting Wu, Haishan Shi, Lin Chen, Minhao Yan
Journal: Materials Science and Engineering: C (2017)

Enhanced osteointegration of tantalum-modified titanium implants with micro/nano-topography
Authors: Junyu Shi, Xiaomeng Zhang, Shichong Qiao, Jie Ni, Jiaji Mo, Yingxin Gu, Hongchang Lai
Journal: RSC Advances (2017): 46472--46479

Grafting of Ring-Opened Cyclopropylamine thin films on Silicon (100) Hydride via UV Photoionization
Authors: Joline Tung, Jing Yuan Ching, Yoke Mooi Ng, Lih Shin Tew, Yit Lung Khung
Journal: ACS Applied Materials & Interfaces (2017)

Microtopography Attenuates Endothelial Cell Proliferation by Regulating MicroRNAs
Authors: Dan Wang, Mengya Liu, Shuangying Gu, Yue Zhou, Song Li
Journal: Journal of Biomaterials and Nanobiotechnology (2017): 189--201

Study on the Regulation of Focal Adesions and Cortical Actin by Matrix Nanotopography in 3D Environment
Authors: Jingjing Han, Keng-hui Lin, Lock Yue Chew
Journal: Journal of Physics: Condensed Matter (2017)

The correlation between osteopontin adsorption and cell adhesion to mixed self-assembled monolayers of varying charges and wettability
Authors: Lijing Hao, Tianjie Li, Fan Yang, Naru Zhao, Fuzhai Cui, Xuetao Shi, Chang Du, Yingjun Wang
Journal: Biomaterials Science (2017)


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