CellTrace™ CFSE Cell Proliferation Kit, for flow cytometry

Citations & References (3)

Invitrogen™

CellTrace™ CFSE Cell Proliferation Kit, for flow cytometry

Name: CellTrace™ CFSE Cell Proliferation Kit, for flow cytometry
SKU: C34554

CellTrace™ CFSE Cell Proliferation Kitは、in vitroおよびin vivoで細胞を標識し、色素の希釈を利用してフローサイトメトリーにより細胞を複数の世代にわたり追跡するのに使用します。•優れた性能—明るいシングルピークの染色により詳細を見る

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表示を変更

製品番号（カタログ番号）	対応可能対象
C34554	180 Reactions
C34570	20 Reactions

2 オプション

製品番号（カタログ番号） C34554

価格（JPY）

52,600

Each

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対応可能対象:

180 Reactions

CellTrace™ CFSE Cell Proliferation Kitは、in vitroおよびin vivoで細胞を標識し、色素の希釈を利用してフローサイトメトリーにより細胞を複数の世代にわたり追跡するのに使用します。

•優れた性能—明るいシングルピークの染色により、複数世代の可視化が可能
• 長時間のシグナル安定性—染色後数日間、細胞内に保持
•非毒性—染色は細胞の健康状態に悪影響をおよぼさない
• シンプルかつ堅牢な染色プロトコル

フローサイトメトリー用のすべてのCellTrace™ Cell Proliferation Kitの選択ガイドをご覧ください。

優れた蛍光染色
色素希釈による増殖分析（下図参照）を成功させるには、細胞分裂後を複数回経た後に蛍光標識細胞を自家蛍光と区別できるよう、非常に明るい色素が必要です。CellTrace™ CFSE色素による強力な蛍光染色により、内因性の細胞自家蛍光がそのシグナルに優る前に8世代以上の増殖細胞を可視化できます。一貫性のある均一な染色により、集団内の細胞間の蛍光変動がほとんどないため、複雑なモデリングソフトウェアを必要とせずに、異なる世代を観察できます。

長期的なシグナル保持
細胞の脂質膜を標識する染色とは異なり、CellTrace™ CFSE色素は細胞膜を通過しやすく、細胞内で共有結合します。この場合、安定した保持力のある蛍光色素は、細胞培養環境で数日経過しても一貫したシグナルが得られる仕様になっています。

非毒性
CellTrace™ CFSE色素は、細胞表面および細胞内部のすべての遊離アミンと共有結合し、細胞毒性を示すことはほぼ皆無であり、細胞増殖性や生物学への影響も最小限に抑えます。研究者たちは、増殖因子カクテルによる刺激に反応して、移植可能な造血細胞がin vitroで増殖することを示すために、CFSE SE標識を使用してきました。

シンプルで堅牢な染色プロトコル
CellTrace™ CFSE Cell Proliferation Kitには、乾燥した色素が入った便利なシングルユースバイアルが含まれており、過剰な量の色素を調製することなく小規模な実験が可能です。使用前に、バイアルの内容物を無水DMSOに溶解してストック溶液を調製します。タンパク質フリー培地中の細胞1 mLを染色するのに、通常このストック溶液1 µLを使用します。細胞を室温で緩やかに撹拌して20分間染色する必要があります。完全培地で短時間洗浄して、溶液中に残っている色素を消光させます。

研究用にのみ使用できます。診断用には使用いただけません。

仕様

概要CellTrace™ Cell Proliferation Kit、180 rxns、CFSE、凍結乾燥、蛍光検出法、励起／発光波長492/517 nm、CellTrace™ CFSEラベル／色素、–5℃～30℃で保存、フローサイトメトリーで使用

検出法蛍光

染色剤タイプその他の標識または色素

形状凍結乾燥

フォーマットチューブ

数量1キット

試薬タイプCFSE＆関連化合物

出荷条件室温

溶解性DMSO（ジメチルスルホキシド）

対応可能対象180 Reactions

Emission488

使用対象（アプリケーション）増殖アッセイ, フローサイトメトリー

使用対象 (装置)フローサイトメーター

製品ラインCellTrace

製品タイプ細胞増殖キット

Unit SizeEach

組成および保存条件

CellTrace™ CFSE（凍結乾燥粉末）バイアル10本と、DMSO（500 μL）バイアル1本入り。フリーザー（-5～-30度）に保存し、遮光してください。

よくあるご質問（FAQ）

I'm trying to label my cells in suspension and track them over time by labeling with CFDA SE, but after labeling them they will no longer adhere to a surface (whereas unlabeled cells adhere well). I've tried to label at 10 and 20 µM. What is causing this?

This dye will bind to proteins via primary amines. Too high of a concentration can lead to cell toxicity or unintended modification of cell function. The solution is to reduce the concentration and/or staining time, or to go with a non-protein-binding tracking reagent, such as Qtracker cell labeling reagents.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I want to do a cell migration study for around 4 hours and need to fluorescently label the cells with a dye. What do you recommend?

Calcein, AM and FDA (fluorescein diaceate) are examples of some dyes used for this application. Since these dyes are not incorporated or covalently attached to any cellular components, they may have a short retention time as some cell types may actively efflux the dye out of the cells. The CellTracker and CellTrace dyes include either a mild thiol-reactive chloromethyl group or amine-reactive succinnimidyl ester group to allow for covalent binding to cellular components, providing for better retention. As with any reagent, one should empirically determine retention times for the cell type used.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I labeled my cells with Calcein, AM, but when I imaged the next day, there was no fluorescence from Calcein. Why?

Calcein, AM is a good choice for cell tracking and as a general cytoplasmic stain. However, it doesn't bind to anything and may be actively pumped out of the cells within a couple hours, which is likely what happened. The retention of Calcein within live cells is dependent upon the inherent properties of the cell type and culture conditions.

For long-term imaging, you may wish to consider a reactive cytoplasmic stains such as CFDA, SE or the CellTracker and CellTrace dyes.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What is the shelf life of CellTrace CFSE Cell Proliferation Kit (Cat. No. C34554, C34570)?

CellTrace CFSE Cell Proliferation Kit is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended (in freezer (-5 to -30 degrees C) and protected from light). Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf).

For staining with CellTrace CFSE Cell Proliferation Kit, for flow cytometry (Cat. No. C34554 or C34570), up to how many cells can I have in 1 mL volume?

The cell concentration can be up to 1 x 10E7 cells/mL.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

View all CellTrace™ CFSE Cell Proliferation Kit, for flow cytometry, 180 reactions kit FAQs

引用および参考文献 (3)

引用および参考文献

Abstract

Optimized staining and proliferation modeling methods for cell division monitoring using cell tracking dyes.

Authors:Tario JD, Humphrey K, Bantly AD, Muirhead KA, Moore JS, Wallace PK,

Journal:J Vis Exp

PubMed ID:23271219

'Fluorescent cell tracking dyes, in combination with flow and image cytometry, are powerful tools with which to study the interactions and fates of different cell types in vitro and in vivo.(1-5) Although there are literally thousands of publications using such dyes, some of the most commonly encountered cell tracking applications ... More

Appraising the suitability of succinimidyl and lipophilic fluorescent dyes to track proliferation in non-quiescent cells by dye dilution.

Authors:Filby A, Begum J, Jalal M, Day W,

Journal:

PubMed ID:25802116

Successful completion of the cell cycle usually results in two identical daughter progeny. This process of generational doubling is termed proliferation and when it occurs in a regulated fashion the benefits range from driving embryonic development to mounting a successful immune response. However when it occurs in a dis-regulated fashion, ... More

TLR4 signaling induced by lipopolysaccharide or paclitaxel regulates tumor survival and chemoresistance in ovarian cancer.

Authors:Szajnik M, Szczepanski MJ, Czystowska M, Elishaev E, Mandapathil M, Nowak-Markwitz E, Spaczynski M, Whiteside TL,

Journal:Oncogene

PubMed ID:19826413

Toll-like receptors (TLRs) expressed on immune cells trigger inflammatory responses. TLRs are also expressed on ovarian cancer (OvCa) cells, but the consequences of signaling by the TLR4/MyD88 pathway in these cells are unclear. Here, TLR4 and MyD88 expression in OvCa tissues (n=20) and cell lines (OVCAR3, SKOV3, AD10, A2780 and ... More

CellTrace™ CFSE Cell Proliferation Kit, for flow cytometry

よくあるご質問（FAQ）

I'm trying to label my cells in suspension and track them over time by labeling with CFDA SE, but after labeling them they will no longer adhere to a surface (whereas unlabeled cells adhere well). I've tried to label at 10 and 20 &micro;M. What is causing this?

I want to do a cell migration study for around 4 hours and need to fluorescently label the cells with a dye. What do you recommend?

I labeled my cells with Calcein, AM, but when I imaged the next day, there was no fluorescence from Calcein. Why?

What is the shelf life of CellTrace CFSE Cell Proliferation Kit (Cat. No. C34554, C34570)?

For staining with CellTrace CFSE Cell Proliferation Kit, for flow cytometry (Cat. No. C34554 or C34570), up to how many cells can I have in 1 mL volume?

引用および参考文献 (3)

I'm trying to label my cells in suspension and track them over time by labeling with CFDA SE, but after labeling them they will no longer adhere to a surface (whereas unlabeled cells adhere well). I've tried to label at 10 and 20 µM. What is causing this?