BAPTA
(Synonyms: 1,2-双(2-氨基苯氧基)-乙烷-N,N,N`,N`-四乙酸) 目录号 : GC17574
BAPTA是一种高钙选择性螯合剂，用于分析钙在不同细胞功能中的作用。
Cas No.:85233-19-8
Sample solution is provided at 25 µL, 10mM.
BAPTA is a highly Ca2+-selective chelator used to dissect the roles of calcium in diverse cellular functions[1-2]. The affinity of BAPTA for Ca2+ can be strengthened or weakened by electron-donating or -withdrawing substituents on the aromatic rings, respectively. The Ca2+ binding to BAPTA is independent of pH within cells[3]. In addition, BAPTA can directly inhibited purified human 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) activity[4].
In vitro, treatment of mechanically injured neurons with BAPTA at 10, 20, or 40μM resulted in a dose-dependent reduction in terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells, with maximal protection observed at 40μM[5]. Treatment of HEK293T cells with increasing concentrations of BAPTA (0.1, 1, and 10mM) progressively elevated the open-channel probability of the endogenous calcium-activated chloride channel ANO6 (TMEM16F)[6]. BAPTA (0-10mM) inhibits phospholipase C (PLC) activity in a dose-dependent manner independently of Ca2+[7].
In vivo, following complete transection spinal cord injury, topical application of 10mM BAPTA to the lesion site in female BALB/c mice resulted in significantly higher body weight and Modified Basso Beattie Bresnahan (mBBB) scores than in vehicle-treated controls[4]. Iontophoresis of 150mM BAPTA into the endolymph of the bobtail skink in vivo caused a downward shift in the frequency of individual spontaneous otoacoustic emission (SOAE) peaks, with recovery requiring more than one hour[8].
References:
[1] Saoudi Y, Rousseau B, Doussière J, et al. Calcium-independent cytoskeleton disassembly induced by BAPTA. Eur J Biochem. 2004;271(15):3255-3264.
[2] Tsien RY. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980;19(11):2396-2404.
[3] Sneyers F, Speelman-Rooms F, Verhelst SHL, Bootman MD, Bultynck G. Cellular effects of BAPTA: Are they only about Ca2+ chelation?. Biochim Biophys Acta Mol Cell Res. 2024;1871(2):119589.
[4] Sneyers F, Kerkhofs M, Speelman-Rooms F, et al. Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cells. Cell Death Dis. 2023;14(9):600.
[5] Kang KR, Kim J, Ryu B, et al. BAPTA, a calcium chelator, neuroprotects injured neurons in vitro and promotes motor recovery after spinal cord transection in vivo. CNS Neurosci Ther. 2021;27(8):919-929.
[6] Kolesnikov DO, Nomerovskaya MA, Grigorieva ER, et al. Calcium chelation independent effects of BAPTA on endogenous ANO6 channels in HEK293T cells. Biochem Biophys Res Commun. 2024;693:149378.
[7] Hardie RC. Inhibition of phospholipase C activity in Drosophila photoreceptors by 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid (BAPTA) and di-bromo BAPTA. Cell Calcium. 2005;38(6):547-556.
[8] Manley GA, Kirk DL. BAPTA induces frequency shifts in vivo of spontaneous otoacoustic emissions of the bobtail lizard. Audiol Neurootol. 2005;10(5):248-257.
BAPTA是一种高钙选择性螯合剂，用于分析钙在不同细胞功能中的作用[1-2]。BAPTA芳环上取代基的供电子或吸电子性质可分别增强或削弱BAPTA对Ca2+的亲和力。胞内pH变化并不影响BAPTA与Ca2+的结合[3]。此外，BAPTA可以直接抑制纯化的人6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3（PFKFB3）活性[4]。
在体外，用10、20或40μM的BAPTA处理机械损伤的神经元，可剂量依赖性地减少TUNEL阳性细胞，其中40μM时保护作用最强[5]。在HEK293T细胞中，BAPTA浓度自0.1mM递增至10mM，可逐步升高内源性钙激活氯通道ANO6（TMEM16F）的开放概率[6]。0-10mM的BAPTA能以Ca2+非依赖方式剂量依赖性地抑制磷脂酶C（PLC）活性[7]。
在体内，雌性BALB/c小鼠脊髓完全横断后，局部涂抹10mM的BAPTA会使小鼠体重和Modified Basso Beattie Bresnahan（mBBB）评分均显著优于溶剂对照[4]。将150mM的BAPTA离子电渗入短尾石龙子体内内淋巴，可致单个自发耳声发射（SOAE）峰频率向下偏移，恢复时间超过1小时[8]。
| Kinase experiment [1]: | |
Preparation Method | Measured the kinase activity of the recombinant, purified human 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) enzyme using a cell-free biochemical assay based on measuring the production of ADP in the presence of different concentrations of BAPTA (ranging from 1µM to 1mM). |
Reaction Conditions | 1µM-1mM |
Applications | BAPTA directly inhibited PFKFB3 activity. |
| Cell experiment [2]: | |
Cell lines | Neurons |
Preparation Method | For physically damaging neurons, 100mg of glass beads were mixed with differentiated neurons sunk with DPBS. The plates were gently rocked three times to let the beads roll over the cells. DPBS and the glass beads were removed by suction. Before the experiment, BAPTA was prepared by diluting in DMEM/F12 at final concentrations of 10μM, 20μM, and 40μM. Each of the prepared BAPTA (BAPTA group) or DPBS (vehicle control group) was gently treated on the cell. Then, the plate was incubated at 37°C for 30 minutes so that BAPTA could protect the damaged cells. During this process, the sham group was not damaged, and no specific substances were treated. Cells were detached by treatment with TrypLE and incubating the plate at 37°C for 5 minutes. Finally, the cells were collected in DMEM/F12. |
Reaction Conditions | 10, 20, and 40μM; 30min |
Applications | In the BAPTA-treated group, 5.3% of the cells had detached from the plate, versus 14.5% in the vehicle control group. Under SEM, neurons in the vehicle control group displayed apoptotic traits. The number of Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells was significantly decreased in a dose-dependent manner, and at 40μM, BAPTA was maximally effective. |
| Animal experiment [2]: | |
Animal models | BALB/c female mice |
Preparation Method | Six-week-old BALB/c female mice were anesthetized using Zoletil (1:1 composite of tiletamine and zolazepam) and xylazine (3:1 ratio, 1mL/kg). Spinal laminectomy was performed at the T9 level. For treatment, 20μL of 10mM BAPTA diluted in DPBS and DPBS were prepared. The BAPTA or DPBS was treated in drops on the full-transection spot with a pipette and was arranged into the BAPTA group and vehicle control group, respectively. The muscle and fascia were sutured, and the skin was closed. Following the surgery, additional water and crushed food were provided for the first 14 days after Spinal cord injury (SCI). Mice had their bladders expressed manually three times a day. Animals in a sham group were subjected to the same surgical procedures except that spinal cords were not injured. |
Dosage form | 20μL, 10mM; topical application |
Applications | The Modified Basso Beattie Bresnahan (mBBB) scores in the BAPTA group were significantly higher than the scores of the vehicle control from the 3rd day following SCI. The bodyweight of the BAPTA group was significantly higher than that of the vehicle control from the 23th day following SCI. |
References: | |
| Cas No. | 85233-19-8 | SDF | |
| 别名 | 1,2-双(2-氨基苯氧基)-乙烷-N,N,N`,N`-四乙酸 | ||
| 化学名 | 2,2',2'',2'''-(((ethane-1,2-diylbis(oxy))bis(2,1-phenylene))bis(azanetriyl))tetraacetic acid | ||
| Canonical SMILES | OC(CN(CC(O)=O)C1=CC=CC=C1OCCOC2=CC=CC=C2N(CC(O)=O)CC(O)=O)=O | ||
| 分子式 | C22H24N2O10 | 分子量 | 476.23 |
| 溶解度 | DMF: 20 mg/ml,DMSO: 20 mg/ml,DMSO:PBS (pH 7.2) (1:1): 0.5 mg/ml | 储存条件 | Store at -20°C |
| General tips | 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 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.0998 mL | 10.4991 mL | 20.9983 mL |
| 5 mM | 0.42 mL | 2.0998 mL | 4.1997 mL |
| 10 mM | 0.21 mL | 1.0499 mL | 2.0998 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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