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ReadiLink?AF488抗體快速標記試劑盒

貨號1275存儲條件 Multiple
規格2x50 ug Labelings價格1800
Ex (nm)499Em (nm)520
分子量溶劑
產品詳細介紹


簡要概述

Readilink™蛋白標記技術是制備用于熒光成像和流式細胞儀應用的熒光抗體綴合物的最強大,最方便的工具之一。ReadiLink™AF488抗體快速標記試劑盒為制造AlexaFluor®488標記的抗體綴合物提供了最便捷的工具(AlexaFluor®是ThermoFisher Scientific的商標)。此ReadiLink™試劑盒中使用的AF488染料相當穩定,并且對蛋白質氨基具有良好的反應性和選擇性。該試劑盒具有用于標記?2x50 ug抗體的所有基本組分。試劑盒中提供的兩個AF488染料小瓶中的每一個均經過優化,可標記約50 µg抗體。制備的AF488綴合物(與試劑盒一起使用)無需進一步純化即可直接用于熒光成像和流式細胞術。它提供了制備AF488標記抗體的最便捷方法。

點擊查看光譜



產品說明書

樣品分析方案 

工作溶液配制

蛋白質工作溶液(溶液A)
為了標記50 µg蛋白質(假設目標蛋白質濃度為1 mg / mL),請將5 µL(反應總體積的10%)反應緩沖液(組分B)與50 µL目標蛋白質溶液混合。
注意:如果蛋白質濃度不同,請相應地調整蛋白質體積,以使?50 µg蛋白質可用于標記反應。
注意:要標記100 µg蛋白質(假設目標蛋白質濃度為1 mg / mL),請將10 µL(反應總體積的10%)反應緩沖液(組分B)與100 µL目標蛋白質溶液混合。
注意:蛋白質應溶于1X磷酸鹽緩沖鹽水(PBS),pH 7.2-7.4;如果蛋白質溶解在甘氨酸緩沖液中,則必須使用1X PBS(pH 7.2-7.4)進行透析,或使用10 kDa的Amicon Ultra-0.5,Ultracel-10膜(密理博公司的產品目錄UFC501008)去除游離的胺或銨鹽(例如硫酸銨和乙酸銨)。
注意:抗體的純度非常重要。
注意:為獲得最佳標記效率,建議最終蛋白質濃度范圍為1-2 mg / mL。
重要:在打開之前,將所有組分加熱并短暫離心小瓶,并在開始綴合之前立即準備所需的溶液。以下方案僅供參考。

 

操作步驟

進行綴合反應
  1. 將蛋白質工作溶液(溶液A)添加到一個小瓶標記染料(AF488-組分A)中,并通過將小瓶渦旋幾秒鐘將其充分混合。
    注意:如果要標記100 µg的蛋白質,請使用兩個小瓶(組分A),將100 µg的蛋白質分成2 x 50 µg的蛋白質,并使每個50 µg的蛋白質與一小瓶的標記染料反應。然后合并兩個小瓶,用于下一步。
  2. 將綴合反應混合物在室溫下放置30-60分鐘。
    注意:如果需要,可以旋轉或搖動綴合反應混合物更長的時間。 

 

停止綴合反應
  1. 將5 µL(對于50 µg蛋白質)或10 µL(對于100 µg蛋白質)添加到結合??反應混合物中,占TQ™染色猝滅緩沖液(組分C)總反應體積的10%;混合均勻。
  2. 在室溫下孵育10分鐘。標記的蛋白(抗體)現在可以使用了。 

蛋白質結合物的儲存
蛋白質綴合物應在載體蛋白(例如0.1%牛血清白蛋白)存在下以> 0.5 mg / mL的濃度存儲。為了更長的存儲時間,可以將蛋白質結合物凍干或分成單份使用,并存儲在≤–20°C下。
 
圖示

圖1.分別用和不用小鼠抗微管蛋白染色的HeLa細胞,然后用ReadiLink™Rapid AF488抗體標記試劑盒(Cat#1275)制備的Alpha Fluor™488山羊抗小鼠IgG結合物的免疫熒光分析。

 

參考文獻

A Fluorescent Probe to Unravel Functional Features of Cannabinoid Receptor CB1 in Human Blood and Tonsil Immune System Cells.
Authors: Martín-Fontecha, Mar and Angelina, Alba and Rückert, Beate and Rueda-Zubiaurre, Ainoa and Martín-Cruz, Leticia and van de Veen, Willem and Akdis, Mübeccel and Ortega-Gutiérrez, Silvia and López-Rodríguez, María Luz and Akdis, Cezmi A and Palomares, Oscar
Journal: Bioconjugate chemistry (2018): 382-389

N-terminal and central segments of the type 1 ryanodine receptor mediate its interaction with FK506-binding proteins.
Authors: Girgenrath, Tanya and Mahalingam, Mohana and Svensson, Bengt and Nitu, Florentin R and Cornea, Razvan L and Fessenden, James D
Journal: The Journal of biological chemistry (2013): 16073-84

SNAP-tag based agents for preclinical in vitro imaging in malignant diseases.
Authors: Amoury, Manal and Blume, Tobias and Brehm, Hannes and Niesen, Judith and Tenhaef, Niklas and Barth, Stefan and Gattenlöhner, Stefan and Helfrich, Wijnand and Fitting, Jenny and Nachreiner, Thomas and Pardo, Alessa
Journal: Current pharmaceutical design (2013): 5429-36

Transient focal membrane deformation induced by arginine-rich peptides leads to their direct penetration into cells.
Authors: Hirose, Hisaaki and Takeuchi, Toshihide and Osakada, Hiroko and Pujals, Sílvia and Katayama, Sayaka and Nakase, Ikuhiko and Kobayashi, Shouhei and Haraguchi, Tokuko and Futaki, Shiroh
Journal: Molecular therapy : the journal of the American Society of Gene Therapy (2012): 984-93

Chemical visualization of an attractant peptide, LURE.
Authors: Goto, Hiroaki and Okuda, Satohiro and Mizukami, Akane and Mori, Hitoshi and Sasaki, Narie and Kurihara, Daisuke and Higashiyama, Tetsuya
Journal: Plant & cell physiology (2011): 49-58

Multifunctional surface modification of gold-stabilized nanoparticles by bioorthogonal reactions.
Authors: Li, Xiuru and Guo, Jun and Asong, Jinkeng and Wolfert, Margreet A and Boons, Geert-Jan
Journal: Journal of the American Chemical Society (2011): 11147-53

Simultaneous detection of changes in protein expression and oxidative modification as a function of age and APOE genotype.
Authors: DeKroon, Robert M and Osorio, Cristina and Robinette, Jennifer B and Mocanu, Mihaela and Winnik, Witold M and Alzate, Oscar
Journal: Journal of proteome research (2011): 1632-44

Site-specific labeling of proteins for single-molecule FRET measurements using genetically encoded ketone functionalities.
Authors: Lemke, Edward A
Journal: Methods in molecular biology (Clifton, N.J.) (2011): 3-15

Transmission electron microscopy characterization of fluorescently labelled amyloid β 1-40 and α-synuclein aggregates.
Authors: Anderson, Valerie L and Webb, Watt W
Journal: BMC biotechnology (2011): 125

Distribution of nerve endings in the human dorsal radiocarpal ligament.
Authors: Tomita, Kazunari and Berger, Evelyn J and Berger, Richard A and Kraisarin, Jirachart and An, Kai-Nan
Journal: The Journal of hand surgery (2007): 466-73



說明書
ReadiLink?AF488抗體快速標記試劑盒.pdf


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