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Brefeldin A

产品编号 T6062Cas号 20350-15-6
别名 BFA, Ascotoxin, 布雷非德菌素 A, Cyanein, Decumbin

Brefeldin A (Cyanein) 属于大环内酯类抗生素,是一种 ATPase 抑制剂 (IC50=0.2 μM)。Brefeldin A 可以诱导肿瘤细胞分化和凋亡,也具有抑制自噬的活性。

Brefeldin A
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Brefeldin A

纯度: 99.89%
产品编号 T6062 别名 BFA, Ascotoxin, 布雷非德菌素 A, Cyanein, DecumbinCas号 20350-15-6

Brefeldin A (Cyanein) 属于大环内酯类抗生素,是一种 ATPase 抑制剂 (IC50=0.2 μM)。Brefeldin A 可以诱导肿瘤细胞分化和凋亡,也具有抑制自噬的活性。

规格价格库存数量
5 mg¥ 414现货
10 mg¥ 698现货
50 mg¥ 2,470现货
100 mg¥ 3,859期货
1 mL x 10 mM (in DMSO)¥ 414现货
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纯度:99.89%
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产品介绍

生物活性
产品描述
Brefeldin A (Cyanein) belongs to the class of macrolide antibiotics and is an ATPase inhibitor (IC50=0.2 μM). Brefeldin A can induce tumor cell differentiation and apoptosis, and also possesses autophagy inhibitory activity.
靶点活性
ATPase (HCT 116 cells):0.2 μM
体外活性
方法:肿瘤细胞 HL60 、K562 和 HT-29 用 Brefeldin A (2 μM) 处理 72 h,使用 DNA filter elution assay 检测 DNA 片段。
结果:Brefeldin A 以不同的动力学诱导 DNA 断裂。HL60 细胞在 15 h 内观察到完整的 DNA 片段,而 K562 和 HT-29 细胞则需要 48-72 h。[1]
方法:人乳腺癌细胞 MDA-MB-231 用 Brefeldin A (0.05-1 μg/mL) 处理 24 h,使用 Western Blot 方法检测靶点蛋白表达水平。
结果:PARP 切割是细胞死亡的标志性事件,可以在 Brefeldin A 处理的悬浮 MDA-MB-231 细胞中检测到。[2]
体内活性
方法:为检测体内抗肿瘤活性,将 Brefeldin A (15 mg/kg in 10% ricinus oil+5% DMSO+10% ethanol+75% physiologic saline) 腹腔注射给携带人结直肠癌肿瘤 HCT116 的 BALB/c 小鼠,每天一次,持续四周。
结果:Brefeldin A 在体内抑制 HCT116 肿瘤生长。[3]
方法:为检测体内抗肿瘤活性,将 Brefeldin A (16-64 mg/kg in distilled water containing 0.05% Tween 80) 腹腔注射给携带肿瘤 LOX IMVI 的 Athymic NCr nu/nu 小鼠,每天两次,持续五天。
结果:Brefeldin A 在体内显示出抗肿瘤活性,小鼠寿命增加 65%-100%,第 60 天幸存者增加 17%-50%。[4]
激酶实验
ELISA-based active site binding assay: Samples (lysed cells or tissue homogenates) are treated for 1 h at room temperature with the biotinylated active site probe PR-584 (5-15 μM). Samples are denatured by addition of SDS (0.9% final) and heating to 100 °C for 5 min. The denatured samples are transferred to a 96-well or 384-well filter plat, mixed with streptavidin-sepharose beads (2.5-5 μL packed beads/well), and incubated for 1 h at room temperature on a plate shaker. The beads are washed 5 times with 100-200 μL /well of ELISA buffer (PBS, 1% bovine serum albumin, 0.1% Tween-20) by vacuum filtration. The beads are incubated overnight at 4 °C on a plate shaker with the following antibodies recognizing the six catalytic subunits diluted into ELISA buffer: β5, β1, and β2 diluted 1:3000, LMP7 and LMP2 diluted 1:5000, and MECL-1 diluted 1:1000. The beads are washed 5 times with 100-200 μL /well of ELISA buffer and incubated with HRP-conjugated secondary antibody diluted 1:5000 in ELISA buffer and incubated 2 h at room temperature on a plate shaker. The beads are washed 5 times with 100-200 μL /well of ELISA buffer and developed for chemiluminsecence signal using the supersignal ELISA pico substrate following the manufacturer's instructions. Luminescence is measured on a plate reader and converted to ng of proteasome or μg/ml of lysate by comparison with 20S proteasome or untreated cell lysate standard curves. For proteasome inhibitor studies, active site probe binding values are expressed as the percent of binding relative to DMSO treated cells.
细胞实验
HF1A3, HF4.9 cell viability upon the treatments is tested using double staining of cells with YO-PRO 1/PI and SYTO16/PI probes. To access cell proliferation, cells are treated with 0–100 ng/mL Brefeldin A in complete medium for 20 hours before adding 1 μCi/mL [methyl-3H]-thymidine for additional 4 hours at 37 °C. The incorporated radioactive thymidine is quantified by scintillation counting with Microbeta counter. To examine long-term effects of Brefeldin A treatment, cells are seeded at initial concentration 105 cells/mL and treated with 0-75 ng/mL Brefeldin A for up to 5 days. At the time indicated, a sample of cells is removed and viable cell number is assessed by standard Trypan Blue exclusion assay.(Only for Reference)
别名BFA, Ascotoxin, 布雷非德菌素 A, Cyanein, Decumbin
化学信息
分子量280.36
分子式C16H24O4
CAS No.20350-15-6
储存&溶解度
存储store at low temperature | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice.
溶解度信息
Ethanol: 2.8 mg/mL (10 mM)
DMSO: 14 mg/mL (50 mM)
溶液配制表
Ethanol/DMSO
1mg5mg10mg50mg
1 mM3.5668 mL17.8342 mL35.6684 mL178.3421 mL
5 mM0.7134 mL3.5668 mL7.1337 mL35.6684 mL
10 mM0.3567 mL1.7834 mL3.5668 mL17.8342 mL
DMSO
1mg5mg10mg50mg
20 mM0.1783 mL0.8917 mL1.7834 mL8.9171 mL
50 mM0.0713 mL0.3567 mL0.7134 mL3.5668 mL

计算器

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  • 配液 计算器
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体内实验配液计算器

请在以下方框中输入您的动物实验信息后点击计算,可以得到母液配置方法和体内配方的制备方法:
TargetMol | Animal experiments比如您的给药剂量是 10 mg/kg ,每只动物体重 20 g ,给药体积 100 μLTargetMol | Animal experiments 一共给药动物 10 只 ,您使用的配方为 5% TargetMol | reagent DMSO+ 30%PEG300+ 5%Tween 80 + 60% ddH2O. 那么您的工作液浓度为 2 mg/mL
母液配置方法: 2 mg 药物溶于 50 μLDMSOTargetMol | reagent ( 母液浓度为 40 mg/mL ), 如您需要配置的浓度超过该产品的溶解度,请先与我们联系。
体内配方的制备方法:50μLDMSOTargetMol | reagent 母液,添加 300 μLPEG300TargetMol | reagent 混匀澄清,再加 50μLTween 80, 混匀澄清,再加 600μLddH2OTargetMol | reagent 混匀澄清

以上为“体内实验配液计算器”的使用方法举例,并不是具体某个化合物的推荐配制方式,请根据您的实验动物和给药方式选择适当的溶解方案。

1 请输入动物实验的基本信息
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% DMSO
%
%Tween 80
%ddH2O

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