Bafilomycin A1 (Baf-A1)

Catalog No.S1413 Batch:S141309

Print

Technical Data

Formula

C35H58O9
Molecular Weight 622.83 CAS No. 88899-55-2
Solubility (25°C)* In vitro DMSO 100 mg/mL (160.55 mM)
Water Insoluble
Ethanol Insoluble
In vivo (Add solvents to the product individually and in order)
Clear solution
5% DMSO 95% Corn oil
1.65mg/ml Taking the 1 mL working solution as an example, add 50 μL of 33 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results. 
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O
5.0mg/ml Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
* <1 mg/ml means slightly soluble or 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.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description Bafilomycin A1(Baf-A1) is a vacuolar H+-ATPase inhibitor with IC50 of 0.44 nM. Bafilomycin A1 is found to inhibit autophagy while induces apoptosis.
Targets
H+-ATPase [1]
(Cell-free assay)
0.44 nM
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]
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 (from reference)

Kinase Assay:

[2]
  • 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.
Cell Assay:

[4]
  • Cell lines

    HeLa cells

  • Concentrations

    ~20 nM

  • Incubation Time

    20 hours

  • Method

    H. pylori bacteria extract is treated with inhibitors, before addition to HeLa cells, as follows: DCCD 10 mM for 1 hour at 30 ºC; NBD-CI 100 μM for 1 hour at 30 ºC and the reaction is blocked with glycine 10 mM final concentration; NEM 275 μM for 1 hour at 30 ºC and the reaction is blocked by addition of β-mercaptoethanol 275 mM; Mg-ATP 14 μM for 1 hour at 0 ºC; 100 μM KNO3 and 14 μM Mg-ATP for 1 hour at 30 μM; NaCO3 100 μM, pH 11 for 1 hour at 0 ºC. The bacterial extract is then added to cell with a 40-fold dilution at a final concentration of 0.65 mg/mL. Controls are HeLa cells incubated with untreated bacterial extracts and cells treated with inhibitor Bafilomycin A1 under the same conditions as bacterial extracts, at the same concentrations or after a 40-fold dilution. The vacuotating activity of the bacterial extracts is assayed.
Animal Study:

[9]
  • Animal Models

    Young (approximately 9 days old) Mozambique tilapia, Oreochromis mossambicus

  • Dosages

    10 μM

  • Administration

    --

Customer Product Validation

Data from [Data independently produced by , , Chem Sci, 2017, 8(3):1915-1921]

Data from [Data independently produced by , , Cancer Lett, 2018, 431:85-95]

Data from [Data independently produced by , , J Exp Clin Cancer Res, 2018, 37(1):251]

Data from [Data independently produced by , , EBioMedicine, 2018, 33:242-252]

Selleck's Bafilomycin A1 (Baf-A1) has been cited by 494 publications

Stress response silencing by an E3 ligase mutated in neurodegeneration [ Nature, 2024, 626(8000):874-880] PubMed: 38297121
Stress response silencing by an E3 ligase mutated in neurodegeneration [ Nature, 2024, 626(8000):874-880] PubMed: 38297121
The Fanconi anemia pathway induces chromothripsis and ecDNA-driven cancer drug resistance [ Cell, 2024, 187(21):6055-6070.e22] PubMed: 39181133
Co-opting templated aggregation to degrade pathogenic tau assemblies and improve motor function [ Cell, 2024, 187(21):5967-5980.e17] PubMed: 39276772
ANKFY1 bridges ATG2A-mediated lipid transfer from endosomes to phagophores [ Cell Discov, 2024, 10(1):43] PubMed: 38622126
A TNIP1-driven systemic autoimmune disorder with elevated IgG4 [ Nat Immunol, 2024, 10.1038/s41590-024-01902-0] PubMed: 39060650
Sensing steroid hormone 17α-hydroxypregnenolone by GPR56 enables protection from ferroptosis-induced liver injury [ Cell Metab, 2024, S1550-4131(24)00371-1] PubMed: 39389061
PRDX6 augments selenium utilization to limit iron toxicity and ferroptosis [ Nat Struct Mol Biol, 2024, 10.1038/s41594-024-01329-z] PubMed: 38867112
SLC13A3 is a major effector downstream of activated β-catenin in liver cancer pathogenesis [ Nat Commun, 2024, 15(1):7522] PubMed: 39215042
Autophagy regulates the maturation of hematopoietic precursors in the embryo [ Nat Commun, 2024, 15(1):2255] PubMed: 38490977

RETURN POLICY
Selleck Chemical’s Unconditional Return Policy ensures a smooth online shopping experience for our customers. If you are in any way unsatisfied with your purchase, you may return any item(s) within 7 days of receiving it. In the event of product quality issues, either protocol related or product related problems, you may return any item(s) within 365 days from the original purchase date. Please follow the instructions below when returning products.

SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.