Brefeldin A
作者：齊岳
發(fā)布時(shí)間：2023-09-12
09:13:46
點(diǎn)擊數(shù)：
產(chǎn)品名稱：Brefeldin A
產(chǎn)品描述：
| 植物來(lái)源 | 天然產(chǎn)物 > 其他（植物來(lái)源） > 其他 |
| 產(chǎn)品描述 | Brefeldin A , a Penicillium brefeldianum metabolite, which is a macrocyclic lactone exhibiting a wide range of antibiotic activity. It is also an ATPase inhibitor for protein transport in HCT 116 cells(IC50=0.2 μM), inducing Y cell differentiation and apoptosis. |
| 靶點(diǎn)活性 | ATPase (HCT 116):0.2 μM |
| 體內(nèi)活性 | 在HF4.9 和 HF28RA 細(xì)胞中, Brefeldin A（25 ng/mL）完全抑制細(xì)胞生長(zhǎng)。在HF1A3 細(xì)胞中,Brefeldin A (75 ng/mL) 完全抑制細(xì)胞生長(zhǎng)。 |
| 激酶實(shí)驗(yàn) | 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. |
| 細(xì)胞實(shí)驗(yàn) | 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) |
| 別名 | Ascotoxin, Cyanein, BFA, 布雷非德菌素 A, Decumbin |
| 分子量 | 280.36 |
| 分子式 | C16H24O4 |
| CAS No. | 20350-15-6 |
存儲(chǔ)
Powder: -20°C for 3 years | In solvent: -80°C for 2 years
溶解度
Ethanol: 2.8 mg/mL (10 mM)
DMSO: 14 mg/mL (50 mM)
( < 1 mg/mL refers to the product slightly soluble or insoluble )
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