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鬼筆環肽-熒光素標記

貨號23101存儲條件 在零下15度以下保存, 避免光照
規格300 Tests價格1896
Ex (nm)498Em (nm)517
分子量~1100溶劑DMSO
產品詳細介紹


簡要概述

產品外觀形態:液體(注:鬼筆環肽系列產品有固體和液體兩種,液體為DMSO溶解后的產品,則不需再溶解。)

這種綠色熒光鬼筆環肽綴合物(相當于FITC標記的鬼筆環肽)會選擇性結合F-肌動蛋白。鬼筆環肽衍生物經常以納摩爾濃度使用,常在甲醛固定和透化的組織切片、細胞培養或無細胞實驗中用于標記、鑒定和定量F-肌動蛋白的便捷探針。鬼筆環肽與肌動蛋白絲的結合比與肌動蛋白單體的結合要緊密得多,從而導致肌動蛋白亞基從絲端解離的速率常數降低,從而通過防止絲解聚而基本穩定了肌動蛋白絲。而且,發現鬼筆環肽抑制F-肌動蛋白的ATP水解活性。鬼筆環肽在細胞中不同濃度下的功能不同。當以低濃度引入細胞質時,鬼筆環肽將聚合度較低的胞質肌動蛋白和纖維蛋白吸收到聚集的肌動蛋白聚合物的穩定“島”中,但它不會干擾應力纖維,即厚的微絲束。鬼筆環肽的性質是研究F-肌動蛋白在細胞中分布的有效工具,方法是用熒光類似物標記鬼筆環肽并將其用于染色肌動蛋白絲以進行光學顯微鏡觀察。鬼筆環肽的熒光衍生物已被證明在定位活細胞或固定細胞中的肌動蛋白絲以及體外觀察單個肌動蛋白絲方面非常有用。熒光鬼筆環肽衍生物已被用作高分辨率研究肌動蛋白網絡的重要工具。鬼筆環肽-熒光素標記探針是AAT Bioquest為多色成像應用提供的一種熒光鬼筆環肽衍生物。

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產品說明書

樣品分析方案

概述

在微孔板孔中制備樣品

從板中的樣品中取出液體

添加鬼筆環肽-熒光素標記溶液(100μL/孔)

在室溫下染色細胞20至90分鐘

清洗細胞在顯微鏡下觀察樣品

注意:將小瓶加熱至室溫并在打開前短暫離心。

 

操作步驟

1.制備1X鬼筆環肽綴合物工作溶液:加入1μL1000X鬼筆環肽綴合物DMSO溶液至1mL含1%BSA的PBS中。

注意1:鬼筆環肽綴合物未使用的1000X DMSO儲備溶液應等分并儲存在-20℃,避光。

注意2:不同的細胞類型可能染色不同。鬼筆環肽綴合物工作溶液的濃度的制備需要根據實際而定。

2.染色細胞:

2.1執行甲醛固定,在室溫下將含3.0-4.0%甲醛的細胞在PBS中孵育10-30分鐘。

注意:避免使用任何含甲醇的固定劑,因為甲醇會在固定過程中破壞肌動蛋白。 優選的固定劑是不含甲醇的甲醛。

2.2用PBS沖洗固定細胞2-3次。

2.3可選:在PBS中加入0.1%Triton X-100固定細胞(步驟2.2)3至5分鐘,以增加滲透性。 用PBS沖洗細胞2-3次。

2.4將100μL/孔(96孔板)鬼筆環肽綴合物工作溶液(來自步驟1)加入固定的細胞中,并在室溫下染色細胞20至90分鐘。

2.5在加上蓋玻片之前,用PBS輕輕沖洗細胞2至3次以除去過量的鬼筆環肽綴合物,然后在顯微鏡下進行,密封和成像。

 

試劑應用文獻

Mussel-inspired conductive nanofibrous membranes repair myocardial infarction by enhancing cardiac function and revascularization

Authors: Yutong He, Genlan Ye, Chen Song, Chuangkun Li, Weirong Xiong, Lei Yu, Xiaozhong Qiu, Leyu Wang
Journal: Theranostics. 2018; 8(18): 5159–5177.

 

參考文獻

Antimicrobial Titanium Surface via Click-Immobilization of Peptide and Its in Vitro/Vivo Activity
Authors: Junjian Chen, Yuchen Zhu, Menghua Xiong, Guansong Hu, Jiezhao Zhan, Tianjie Li, Lin Wang, Yingjun Wang
Journal: ACS Biomaterials Science & Engineering (2018)

Mussel-inspired conductive nanofibrous membranes repair myocardial infarction by enhancing cardiac function and revascularization
Authors: Yutong He, Genlan Ye, Chen Song, Chuangkun Li, Weirong Xiong, Lei Yu, Xiaozhong Qiu, Leyu Wang
Journal: Theranostics (2018): 5159

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

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

Paxillin facilitates timely neurite initiation on soft-substrate environments by interacting with the endocytic machinery
Authors: Ting-Ya Chang, Chen Chen, Min Lee, Ya-Chu Chang, Chi-Huan Lu, Shao-Tzu Lu, De-Yao Wang, Aijun Wang, Chin-Lin Guo, Pei-Lin Cheng
Journal: eLife (2017): e31101



說明書
鬼筆環肽-熒光素標記.pdf


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