الصفحة الرئيسية>>Natural Products>>Calcein-AM (Calcein acetoxymethyl ester)

Calcein-AM (Calcein acetoxymethyl ester) (Synonyms: Calcein Acetoxymethyl ester, NSC 689290)

رقم الكتالوجGC34061

كالسين-أم (Calcein-AM) هو صبغة فلورية قابلة للتخريش تستخدم لتحديد قابلية الخلايا للعيش.

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Calcein-AM (Calcein acetoxymethyl ester) التركيب الكيميائي

Cas No.: 148504-34-1

الحجم السعر المخزون الكميّة
500μg
149٫00
متوفر
1mg
282٫00
متوفر

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Sample solution is provided at 25 µL, 10mM.

Product has been cited by 5 publications

Protocol

Cell experiment [1]:

Cell lines

EPC cells

Preparation Method

Uptake kinetics of EPC cells loaded with 5 µM calcein AM seeded at 1x105 cells well-1 and cultured at 15 °C. Calcein AM uptake was measured as fluorescence intensity (FI).

Reaction Conditions

5 µM at 15 °C, 1-8h

Applications

Uptake kinetics showed that for EPC cells seeded at a density of 1x105 cells well-1 calcein AM labelling increased throughout the 7 h tested.

References:

[1]. Iwanowicz LR, et al. Calcein AM release-based cytotoxic cell assay for fish leucocytes. Fish Shellfish Immunol. 2004 Feb;16(2):127-37.

Background

كالسين-أم هو صبغة حية شديدة الدهونية تدخل بسرعة إلى الخلايا الحية. يتم تحويلها من قبل استيرازات داخل الخلية إلى كالسين الذي ينتج إشارة خضراء مكثفة (530 نانومتر)، ويُحافظ عليه في الخلايا ذات غشاء بلازمائي سليم. [1]

أظهرت التجربة في الأنابيب أن اختبار كالسين-AM يستخدم لتقييم قابلية بقاء خلايا الدم الحمراء البشرية بعد التفاعل (37 درجة مئوية لمدة 3 و 20 ساعة) في وجود Ca2+ (2.5 ملم) و ionophore A23187 (0.5 μM). كان تركيز CAM المثلى هو 0.05 μM لصبغ خلايا المؤثرات عن طريق اختبار 0.05، 0.1، 0.2، و 0.4 μM . باستخدام CAM بتركيز 0.05μm لصبغ PBMCs و NK cells الموسَّع من ثلاثة متطوِّعِینٍ طَیْفِیِّینٍ, أظهَروُا نتائج تشير إلى عدم حُصول انخفاض كافٍ في فعالية NK cell الإنسانية في PBMCs أو NK cells الموسَّع, كذلك صور صُورَەُ کالچین AM's fluorescent signal of lymphocytes x lo5 at a concentration of50μm was close to the saturation level, while the signal emitted by lymphocytes labeled with calcein AM at a concentration of20μm was only slightly lower. كان لدى كالسين-AM نشاط سمِّي ضد خلايا الأورام البشرية (مثل ورم اللمفاويات U-937-GTB) عند تركيزات منخفضة (2.5 مج/مل). أظهَرَت التجربة في الأنابيب أن استخدام اختبار صبغ Calcein-AM / PI double staining بتركیز 2 µM / 4.5 µM لصبغ الماكروفاج وخلايا THP-1 المُخْتَلِطة (5 × 105 خلية / مل) يستخدم لقیاس عدد الخلایای حیة والمیتة کإحصاءٍ لوفاة الخلاية. بالإضافة إلى ذلک، تُزْرع خلايا OA chondrocytes في صحون بها 24 فسيلة (2 × 104 خلية / فسيلە), يُزارع۔ثُـــــﮯ هذهِ الْخَلاِئِین ﻟﺘﻤﺜّﻠھَہ ﮐﺎﻟﺴّ‌چ ﭬیدولین، ويرقدهاأثناء درجات حرارة تصل إلى 37 درجة مئوية في ظروف خالية من الضوء لمدة 30 دقيقة، وتحليلها بعد ذلك تحت المجهر الفلورسنتي.

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.

Chemical Properties

Cas No. 148504-34-1 SDF
المرادفات 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
الذوبان 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.
Shipping Condition Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table

Prepare stock solution
1 mg 5 mg 10 mg
1 mM 1.0052 mL 5.0258 mL 10.0517 mL
5 mM 0.201 mL 1.0052 mL 2.0103 mL
10 mM 0.1005 mL 0.5026 mL 1.0052 mL
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