BAPTA-AM is a well-known membrane permeable Ca2+ chelator. BAPTA-AM inhibits hERG channels, hKv1.3 and hKv1.5 channels in HEK 293 cells with IC50s of 1.3 μM, 1.45 μM and 1.23 μM, respectively[1].
IC50 Target
Ca2+ chelator[1] IC50: 1.3 μM (hERG channel, in HEK 293 cells), 1.45 μM (hKv1.3, in HEK 293 cells), 1.23 μM (hKv1.5, in HEK 293 cells)[1]
体外研究 (In Vitro)
BAPTA-AM inhibits neuronal Ca2+-activated K+ channel currents, and up-regulates the decreased cardiac sodium current (INa) density by chelating intracellular Ca2+[1]. BAPTA-AM (BAPTA/AM), an intracellular calcium chelator, induces delayed necrosis by lipoxygenase-mediated free radicals in mouse cortical cultures. BAPTA-AM prevents free radical-mediated toxicity promote apoptosis in non-neuronal cells and produce a beneficial effect in neuronal cells by protecting neurons from ischemic damage. In addition, it has been suggested that BAPTA-AM induces a late, but not early, increase of intracellular calcium in I-IL-60 neoplastic cells. Mixed cortical cell cultures (DIV 13-16) exposed to 10 μM BAPTA-AM for 24- or 48-hr show moderate (45-70%) neuronal injury as evaluated by increased LDH release into the bathing medium after 24-48-hr. Exposure of cortical cultures to 3-10 μM BAPTA-AM for 48-hr evoke dose-dependent neuronal damage[2].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量
764.68
Formula
C34H40N2O18
CAS 号
126150-97-8
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Tang Q, et al. The membrane permeable calcium chelator BAPTA-AM directly blocks human ether a-go-go-related gene potassium channels stably expressed in HEK 293 cells. Biochem Pharmacol. 2007 Dec 3;74(11):1596-607.
[2]. Wie MB, et al. BAPTA/AM, an intracellular calcium chelator, induces delayed necrosis by lipoxygenase-mediated free radicals in mouse cortical cultures. Prog Neuropsychopharmacol Biol Psychiatry. 2001 Nov;25(8):1641-59.
Cell Assay [1]
Neuronal injury is quantitatively estimated by measuring lactate dehydrogenase (LDH) released from damaged cells into the bathing medium 24- or 48-hr after the 10 μM BAPTA/AM treatment. The morphological findings are confirmed by staining with neuron-specific enolase (NSE) antibody and tryphan blue[1].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献
[1]. Tang Q, et al. The membrane permeable calcium chelator BAPTA-AM directly blocks human ether a-go-go-related gene potassium channels stably expressed in HEK 293 cells. Biochem Pharmacol. 2007 Dec 3;74(11):1596-607.
[2]. Wie MB, et al. BAPTA/AM, an intracellular calcium chelator, induces delayed necrosis by lipoxygenase-mediated free radicals in mouse cortical cultures. Prog Neuropsychopharmacol Biol Psychiatry. 2001 Nov;25(8):1641-59.
Calcein-AM is cell-permeable fluorescent dye used to determine the cell viability.
体外研究 (In Vitro)
The calcein-AM dye used to stain the living cells is shown to have a low spontaneousleakage rate less than 15% in 4 hours at 37°C. Dilutions of targets stained by calcein-AM has a linear relationship with measured fluorescence values. NK cells, LAKs, and CTLs are readily detectable by this microtest. Quantitation of killing and kinetic analysis is readily performed with the test system[1]. Calcein-AM is pH independent, better retained and more photostable. In addition, the high level of intracellular retention of calcein-AM and its low-level release after incorporation exclude possible cell-monolayer labeling and allow its use in a cell-cell interaction assay. Moreover, the bright fluorescence can easily be detected and measured by a microplate fluorescence reader[2]. Calcein-AM is a highly lipophilic vital dye that rapidly enters viable cells, is converted by intracellular esterases to calcein that produces an intense green (530-nm) signal, and is retained by cells with intact plasma membrane. From dying or damaged cells with compromised membrane integrity or from cells expressing multidrug resistance protein (MRP), unhydrolyzed substrates and their fluorescent products are rapidly extruded from cells. The calcein-AM assay has been used to assess the cell viability, cytotoxicity and tp quantitate apoptosis[3].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
体内研究 (In Vivo)
Calcein-AM is found to be suitable for in vivo studies, because it has no deleterious effects on cell function and is, indeed, a marker of cell viability[2].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量
994.86
Formula
C46H46N2O23
CAS 号
148504-34-1
中文名称
钙黄绿素-AM
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Wang XM, et al. A new microcellular cytotoxicity test based on calcein-AM release. Hum Immunol. 1993 Aug;37(4):264-70.
[2]. Braut-Boucher F, et al. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calceinAM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41-51.
[3]. Bratosin D, et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viabilityand aging. Cytometry A. 2005 Jul;66(1):78-84.
Cell Assay [1][2][3]
K562, Daudi, and Chang liver cells are labeled with calcein-AM. Calcein-AM’s excitation and emission wavelengths are 496 nm and 520 nm, respectively. The filter/mirror combination used to detect calcein-AM’s green fluorescence includes the 490-nm excitation and 520-nm emission filters with a dichroic mirror. Differences in the automatic fluorescence readings between the test and control wells determine the results[1]. A simple and sensitive cell-cell adhesion microplate assay is established using the calcein-AM. The procedure involves three steps: the labeling of lymphocytes with an adequate concentration of calcein-AM (20 μM) during a short incubation period (30 min); the adhesion of 2×105 labeled lymphocytes per well to confluent keratinocyte or fibroblast monolayers grown in microtiter plates for 90 min; and, finally, measurement of the fluorescent signal utilizing a new system of cold-light microfluorimetry[2]. Cells are incubated for 15 min in 1 mL of a 1% saponin solution in PBS buffer, pH 7.4, containing 0.05% sodium azide. After saponin permeabilization, 4×105 RBCs in suspension in PBS buffer containing 0.1% saponin and 0.05% sodium azide are incubated (37°C in the dark for 45 min) with calcein-AM to a final concentration of 5 μM, ished three times with the same PBS buffer containing 0.1% saponin and 0.05% sodium azide, and the cell viability is analyzed by flow cytometry[3].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献
[1]. Wang XM, et al. A new microcellular cytotoxicity test based on calcein-AM release. Hum Immunol. 1993 Aug;37(4):264-70.
[2]. Braut-Boucher F, et al. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calceinAM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41-51.
[3]. Bratosin D, et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viabilityand aging. Cytometry A. 2005 Jul;66(1):78-84.
Fluo-4 AM is a fluorescent dye (λex=494 nm, λem=516 nm). Preloaded with Fuo-4 AM, a very bright fluorescence image is observed. In a parallel experiment with fluo-3 AM-loaded cells, the resulting fluorescence image, although clearly discernable in this case, is less bright[1]. Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). 1. Count the cells and take 106 cells from each sample (control and experiment/s). 2. Collect the cells (5 min, 3000×g, 4 °C) and wash once in PBS. 3. Resuspend the cells in 0.5-ml PBS and add 0.5 μl of Fluo-4-AM (1 mM stock) to a final concentration of 1 μM. Incubate at 37 °C for 1 h. 4. Wash the cells three times (2 min, 3000×g) with PBS and finally resuspend in 1-ml PBS. Separate into 2 flow cytometry tubes—0.5 ml in each. 5. Evaluate the staining by flow cytometry and analyze the data by a software such as CellQuest software.
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量
1096.94
Formula
C51H50F2N2O23
CAS 号
273221-67-3
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
-20deg;C, sealed storage, away from moisture and light
*该产品在溶液状态不稳定，建议您现用现配，即刻使用。
参考文献
[1]. Gee KR, et al. Chemical and physiological characterization of fluo-4 Ca(2+)-indicator dyes. Cell Calcium. 2000 Feb;27(2):97-106.
[2]. Fluo-4.
Cell Assay [1]
For measuring fluorescence from cells in suspension, dilutions to 2 to 3×106 cells are made, from cultures of rat basophilic leukemia (RBL) cells. Cells are incubated in suspension in 1 μM dye (including Fluo-4 AM) for 30 min at 37°C. Cell suspensions are then transferred to cuvets for measurements of fluorescence emission intensity by spectrofluorometer[1].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献
[1]. Gee KR, et al. Chemical and physiological characterization of fluo-4 Ca(2+)-indicator dyes. Cell Calcium. 2000 Feb;27(2):97-106.
HKYellow-AM (6/12-mixture) is a fluorogenic probe extracted from patent EP2809666B1, compound 14, which can be used for sensitive and specific detection of peroxynitrite[1].
分子量
1422.40
Formula
C80H78Cl2N4O16
CAS 号
1448821-89-3
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
-20deg;C, sealed storage, away from moisture and light
*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (sealed storage, away from moisture and light)
溶解性数据
In Vitro:;
DMSO : 130 mg/mL (91.39 mM; Need ultrasonic)
配制储备液
浓度溶剂体积质量
1 mg
5 mg
10 mg
1 mM
0.7030 mL
3.5152 mL
7.0304 mL
5 mM
0.1406 mL
0.7030 mL
1.4061 mL
10 mM
0.0703 mL
0.3515 mL
0.7030 mL
*
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。
In Vivo:
请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：
BTC-AM is a low affinity calcium indicator. BTC-AM has substantial calcium-independent fluorescence at all excitation wavelengths. BTC-AM is readily loaded into neurons and is rapidly hydrolysed[1].
分子量
979.91
Formula
C45H45N3O20S
CAS 号
176767-94-5
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
Please store the product under the recommended conditions in the Certificate of Analysis.
参考文献
[1]. Hyrc KL, et al. Neuronal free calcium measurement using BTC/AM, a low affinity calcium indicator. Cell Calcium. 1998;24(3):165-175.
HKYellow-AM (6/12-mixture) is a fluorogenic probe extracted from patent EP2809666B1, compound 14, which can be used for sensitive and specific detection of peroxynitrite[1].
分子量
1422.40
Formula
C80H78Cl2N4O16
CAS 号
1448821-89-3
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
-20deg;C, sealed storage, away from moisture and light
*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (sealed storage, away from moisture and light)
溶解性数据
In Vitro:;
DMSO : 130 mg/mL (91.39 mM; Need ultrasonic)
配制储备液
浓度溶剂体积质量
1 mg
5 mg
10 mg
1 mM
0.7030 mL
3.5152 mL
7.0304 mL
5 mM
0.1406 mL
0.7030 mL
1.4061 mL
10 mM
0.0703 mL
0.3515 mL
0.7030 mL
*
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。
In Vivo:
请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：
Fluo-8 AM is a calcium fluorescent probe that can be loaded into protoplasts to detect calcium in the flesh tissue cells of Malus domestica[1].
分子量
1046.93
Formula
C50H50N2O23
CAS 号
1345980-40-6
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
Please store the product under the recommended conditions in the Certificate of Analysis.
参考文献
[1]. Lina Qiu, et al. Loading calcium fluorescent probes into protoplasts to detect calcium in the flesh tissue cells of Malus domestica. Hortic Res. 2020 Jun 1;7(1):91.
Rhod-2 AM is a fluorescent, mitochondrial probe (λex=552 nm, λem=581 nm).
体外研究 (In Vitro)
A high concentration of Ca2+ in mitochondria is illustrated by punctate labeling in cells, which is consistent with the location of Ca2+ in the mitochondria following fluorescence staining with Rhod-2 AM (Rhod2-AM) at 6 h p.i.. Inhibition of mitochondrial Ca2+ uptake by ruthenium red (RR) or 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS) in IMR5 cells infected with poliovirus (PV) is also illustrated following fluorescence staining with Rhod-2 AM at 6 h p.i.[1].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量
1123.94
Formula
C52H59BrN4O19
CAS 号
145037-81-6
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
-20deg;C, sealed storage, away from moisture and light
*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (sealed storage, away from moisture and light)
参考文献
[1]. Brisac C, et al. Calcium flux between the endoplasmic reticulum and mitochondrion contributes to poliovirus-induced apoptosis. J Virol. 2010 Dec;84(23):12226-35.
Cell Assay [1]
For flow cytofluorometry, cells are harvested, pelleted, and resuspended in ice-cold PBS containing 10 mM glucose, 10% fetal bovine serum (FBS), and 10 μM Rhod-2 AM (Rhod2-AM). Mitochondrial calcium levels are determined by the flow cytofluorometry analysis of aliquots of 4×105 cells. For fluorescence microscopy, IMR5 cells are grown on polylysine-coated (10 μg/mL) slides and stained with 7.5 μM Rhod-2 AM in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS for 2 h before poliovirus (PV) infection. Cells are fixed by incubation for 15 min at 4°C in 4% paraformaldehyde. Cells are washed in PBS, and images are acquired with Zeiss Apotome and Axiovision software[1].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献
[1]. Brisac C, et al. Calcium flux between the endoplasmic reticulum and mitochondrion contributes to poliovirus-induced apoptosis. J Virol. 2010 Dec;84(23):12226-35.
