Bafilomycin A1(Baf-A1)
Catalog No.S1413
Molecular Weight(MW): 622.83
Bafilomycin A1 is a vacuolar H+-ATPase inhibitor with IC50 of 0.44 nM.
Purity & Quality Control
Choose Selective Proton Pump Inhibitors
Biological Activity
| Description | Bafilomycin A1 is a vacuolar H+-ATPase inhibitor with IC50 of 0.44 nM. | ||||||||||||||||||||||||||||||||||||||||||||
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| In vitro |
Bafilomycin A1 is a toxic macrolide antibiotic derived from Streptomyces griseus. Bafilomycin A1 inhibits the short circuit current induced by the outer mantle epithelium (OME). The IC50 and maximum inhibition dose of Bafilomycin A1 are 0.17 μM and 0.5 μM, respectively. [2] In addition, Bilomycin A1 inhibits the acid influx with an IC50 value of 0.4 nM. Bafilomycin A1 inhibits the acidification dose-dependently resulting in a lower quenching, and thus a higher fluorescence. [3] Bafilomycin A1 prevents the vacuolization of Hela cells induced by H. pylori, with an inhibitory concentration giving 50% of maximal (ID50) of 4 nM. Bafilomycin A1 is also very efficient in restoring vacuolated cells to a normal appearance. [4] Bafilomycin A1 also affects the transport of endocytosed material from early to late endocytic compartments. Bafilomycin not only dissipates the low endosomal pH but also blocks transport from early to late endosomes in HeLa cells. [5] Bafilomycin A1 at doses of 0.1-1 μM completely inhibits the acidification of lysosomes revealed by the incubation with acridine orange in BNL CL.2 and A431 cells. [6] When Bafilomycin A1 is added to Hanks' balanced salt solution, endogenous protein degradation is strongly inhibited and numerous autophagosomes accumulated in H-4-II-E cells. Bafilomycin A1 also prevents the appearance of endocytosed HRP in autophagic vacuoles. [7] |
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| Assay |
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| In vivo | Bafilomycin A1 (1 μM and 0.1 μM) completely inhibits the resorptive activity of cultured osteoclasts. [8] Bafilomycin A1 dose-dependently inhibits the rate of Na+ uptake in young tilapia with a Ki of 0.16 μM. [9] |
Protocol
| Kinase Assay: |
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ATPase enzyme activity assays: The ATPase enzyme assay medium contains 6 mM MgSO4, 50 mM HEPES (pH 7.4), 200 mM Na2SO3 (V-ATPase activator), 0.5 mM sodium ortho-vanadate (P-ATPase inhibitor), 0.5 mM sodium azide (F-ATPase inhibitor) and 3 mM Na2ATP. This medium (1.0 mL), with or without the addition of the V-type ATPase inhibitor bafilomycin A1, is incubated with the filtered homogenate (0.1 mL) for 60 minutes at 23–25 °C. The reaction is stopped by the addition of 1 mL of TCA 3%. Spectrometric blanks are prepared as for the enzyme assay with the exception that the tissue sample is added after the acid. Phosphate analysis is accomplished by adding 2 mL of 1-butanol and 0.2 mL molybdate solution (5 g ammonium molybdate, 22 mL H2SO4 to 100 mL). After vortexing for 15 seconds the solution is neutralised with 0.5 mL citrate solution (100 g/500 mL, pH 7.0) and again vortexed for 15 seconds. The solution is then centrifuged (2000 × g; 3 minutes) to separate the butanol phase and the absorbance of this phase is read at 400 nm. Standards of orthophosphate are prepared (0.1 μM–2.0 μM) and treated in the same way as the enzyme activity assays. Enzyme activity is expressed in μmol of orthophosphate liberated per hour and per milligram of protein. V-ATPase activity is considered to be the difference between the total ATPase activity measured in the presence of Na2SO3, sodium orthovanadate and sodium azide and the ATPase activity measured in the presence of these reagents and of the specific V-ATPases inhibitor Bafilomycin A1. |
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References
- [1] al-Fifi ZI, et al. Insect BiochemMol Biol, 1998, 28(4), 201-211.
- [2] Oliveira PF, et al. Comp Biochem Physiol A Mol Integr Physiol, 2004, 139(4), 425-432.
- [3] S?rensen MG, et al. J Bone Miner Res, 2007, 22(10), 1640-1648.
- [4] Papini E, et al. Mol Microbiol, 1993, 7(2), 323-327.
- [5] Bayer N, et al. J Virol, 1998, 72(12), 9645-9655.
- [6] Yoshimori T, et al. J Biol Chem, 1991, 266(26), 17707-17712.
- [7] Yamamoto A, et al. Cell Struct Funct, 1998, 23(1), 33-42.
- [8] Sundquist KT, et al. J Bone Miner Res, 1994, 9(10), 1575-1582.
- [9] Fenwick JC, et al. J Exp Biol, 1999, 202 Pt 24, 3659-3666.
- [10] Kanzawa T, et al. Cell Death Differ, 2004, 11(4), 448-457.
Solubility (25°C)
| In vitro | DMSO | 6 mg/mL (9.63 mM) |
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| Water | Insoluble | |
| Ethanol | Insoluble |
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Information
| Molecular Weight | 622.83 |
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| Formula | C35H58O9 |
| CAS No. | 88899-55-2 |
| Storage | powder |
| in solvent | |
| Synonyms | N/A |
Bio Calculators
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Tech Support
Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.
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Frequently Asked Questions
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Question 1:
How to dissolve it？
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Answer:
S1413 Bafilomycin A1(Baf-A1) is soluble in DMSO at 0.1 mg/ml. Please do not use alcohols as solvent, because this compound will degrade in alcohols.
