Cell Signaling Technology

Product Pathways - Protein Translation

Everolimus #12017

bFRF   ever   everlomis   everloms   everlomus   FKBP12   mTOR   mTORC1   RAD001   VEGF   ver  

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype

Species cross-reactivity is determined by western blot.

Applications Key:

Molecular Characterization

C53H83NO14

Purity

>99%

Structure

Structure

Chemical structure of everolimus.

everolimus的化学结构。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, serum-starved overnight and treated with hIGF-I #8917 (100 ng/ml, 10 min) either with or without Everolimus pretreatment (1 hr) at the indicated concentrations, using Phospho-p70 S6 Kinase (Thr389) Antibody #9205 (upper) or p70 S6 Kinase (49D7) Rabbit mAb #2708 (lower).

Western blot方法分别检测血清饥饿过夜以及hIGF-I #8917 (100 ng/ml, 10 min) 处理的293细胞提取物,上述细胞包含经过和未经所示浓度的Everolimus预处理两种情况,使用的抗体为Phospho-p70 S6 Kinase (Thr389) Antibody #9205 (上图)或p70 S6 Kinase (49D7) Rabbit mAb兔单抗 #2708 (下图)。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, serum-starved overnight and treated with hIGF-I #8917 (100 ng/ml, 10 min) either with or without Everolimus pretreatment (1 hr) at the indicated concentrations, using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb #4858 (upper) or S6 Ribosomal Protein (5G10) Rabbit mAb #2217 (lower).

Western blot方法分别检测血清饥饿过夜以及hIGF-I #8917 (100 ng/ml, 10 min) 处理的HeLa细胞提取物,上述细胞包含经过和未经所示浓度的Everolimus预处理两种情况,使用的抗体为Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb兔单抗 #4858 (上图)或S6 Ribosomal Protein (5G10) Rabbit mAb兔单抗 #2217 (下图)。

Directions for Use

Everolimus is supplied as a lyophilized powder. For a 1 mM stock, reconstitute the 1 mg in 1.04 ml DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used at 10-100 nM either as a pretreatment for 0.5-1 hr prior to treating with a stimulator or is used alone with varying treatment times lasting up to 24 hr. Soluble in DMSO or ethanol at 100 mg/ml; very poorly soluble in water with maximum solubility ~1-10 µM.

Everolimus以冻干粉形式提供。可将1 mg冻干粉重悬于1.04 ml DMSO制成1 mM贮存液。工作浓度和处理时间的长短取决于预期效果,但通常建议使用10-100 nM预处理0.5-1 hr,然后再加刺激物或结合不同的处理时间单独使用,最长可达24hr。可溶解于DMSO或乙醇,浓度为100 mg/ml;水中溶解度极差,最大溶解度为~1-10 µM。

Background

Everolimus, also known as RAD001, is an immunosuppressant analog of rapamycin. Everolimus forms a complex with FKBP12 with an IC50 = ~2 nM in FK506 competitive binding assays (1,2), and this complex then binds to and inhibits mTORC1 (3). Studies have shown that everolimus treatment of cells can lead to the dephosphorylation of mTOR downstream targets (4-7), inhibition of VEGF- and bFRF-stimulated proliferation in HUVE cells (6), and reduction of hypoxia-induced HIF-1 protein levels (7).

Everolimus,也被称为RAD001,是雷帕霉素的一种免疫抑制剂类似物。Everolimus可与FKBP12形成一种在FK506竞争结合试验中IC50 = ~2 nM的复合物(1,2)。该复合物随后可以结合并抑制mTORC1 (3)。研究表明everolimus处理细胞可以导致mTOR下游靶标的去磷酸化(4-7)、抑制VEGF和 bFRF刺激的HUVE增殖(6)以及低氧诱导HIF-1蛋白水平的降低(7)。

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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