Calcein-AM (Calcein acetoxymethyl ester) (Synonyms: Calcein Acetoxymethyl ester, NSC 689290) |
カタログ番号GC34061
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Calcein-AMは、生存細胞に迅速に入る高脂溶性の活性な染料である。
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 148504-34-1
Sample solution is provided at 25 µL, 10mM.
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Calcein-AMは、生存細胞に迅速に入る高脂溶性の活性な染料である。細胞内のエステラーゼによってcalceinに変換され、強い緑色（530 nm）の信号を発生させ、細胞膜が完全な細胞に保持される。
in vitro実験では、Ca2+（2.5 mM）およびionophore A 23187（0.5 μM）の存在下で、ヒト赤血球（RBC）を37°Cで3時間および20時間培養した後の生存率を評価するためにcalcein-AMアッセイが使用された。CAM（Calcein-AM）の最適濃度は0.05 μMであり、0.05、0.1、0.2、および0.4 μMを試験した。0.05 μMのCAMを使用して、3人の通常のボランティアから採取されたPBMCおよび拡張されたNK細胞を染色した結果、細胞毒性に有意な減少は見られず、PBMCまたは拡張されたNK細胞のヒトNK細胞活性にもCAMの染色が有意な影響を与えなかったことが示された。in vitroでは、50μMのcalcein AMで染色された1 x 10^5リンパ球の蛍光シグナルは飽和レベルに近く、20μMのcalcein AMで染色されたリンパ球のシグナルはわずかに低かったと報告されている。Calcein-AMは、低濃度（2.5 μg/ml）でヒトリンパ腫細胞株（U-937-GTBなど）に対して細胞毒性活性を示する。in vitro実験では、混合マクロファージとTHP-1細胞（5x10^5 cells/ml）を用いて、Calcein-AM（2 µM）/propidium iodide（PI）（4.5 µM）の染色アッセイが生存細胞と死細胞の数を定量化するために使用された。さらに、OA軟骨芽細胞は24-wellプレートに2 × 10^4 cells/wellでシードされ、4時間培養された後、光のない条件下で37°Cで30分間Calcein-AM（2 μM）とPI（2 μM）で処理され、その後蛍光顕微鏡で分析された
References:
[1].Bratosin D, et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viability and aging. Cytometry A. 2005 Jul;66(1):78-84.
[2].Jang YY, et al. An improved flow cytometry-based natural killer cytotoxicity assay involving calcein AM staining of effector cells. Ann Clin Lab Sci. 2012 Winter;42(1):42-9.
[3].Braut-Boucher F, et al. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calcein AM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41-51.
[4].Liminga G, et al. Cytotoxic effect of calcein acetoxymethyl ester on human tumor cell lines: drug delivery by intracellular trapping. Anticancer Drugs. 1995 Aug;6(4):578-85.
[5].Xiang N, et al. Gardnerella vaginalis induces NLRP3 inflammasome-mediated pyroptosis in macrophages and THP-1 monocytes. Exp Ther Med. 2021 Oct;22(4):1174.
[6].Zhang L, et al. MicroRNA-140-5p represses chondrocyte pyroptosis and relieves cartilage injury in osteoarthritis by inhibiting cathepsin B/Nod-like receptor protein 3. Bioengineered. 2021 Dec;12(2):9949-9964.
| 細胞実験[1]: | |
細胞株 | EPC細胞 |
準備方法 | EPC細胞を1×10^5細胞/ウェルの密度で播種し、15°Cで培養した。5 µMのカルセインAMでロードしたEPC細胞の取り込み動態を蛍光強度（FI）として測定した。 |
反応条件 | 5 µM、15°C、1〜8時間 |
アプリケーション | 取り込み動態は、1×10^5細胞/ウェルの密度で播種されたEPC細胞において、テストした7時間全体でカルセインAM標識が増加したことを示した。 |
参考文献: | |
| Cas No. | 148504-34-1 | SDF | |
| 同義語 | Calcein Acetoxymethyl ester, NSC 689290 | ||
| Chemical Name | tetrakis(acetoxymethyl) 2,2',2'',2'''-(((3',6'-diacetoxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-4',5'-diyl)bis(methylene))bis(azanetriyl))tetraacetate | ||
| Canonical SMILES | O=C1OC2(C(C=C(CN(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)C(OC(C)=O)=C3)=C3OC4=CC(OC(C)=O)=C(CN(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)C=C24)C5=C1C=CC=C5 | ||
| Formula | C46H46N2O23 | M.Wt | 994.86 |
| 溶解度 | 10 mg/mL in EtOH, MeOH, DMSO, DMF with gentle heating (37°C) for 10 minutes. | Storage | -20°C, protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.0052 mL | 5.0258 mL | 10.0517 mL |
| 5 mM | 201 μL | 1.0052 mL | 2.0103 mL |
| 10 mM | 100.5 μL | 502.6 μL | 1.0052 mL |
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Quality Control & SDS
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- Purity: >98.00% Appearance: A solid
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After trypsinization, the cells were washed twice with PBS followed by incubation of 2 µM calcein-acetoxymethyI ester (GLPBIO) for 30 min at 37 °C.
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