Bafilomycin A1 (Baf-A1)

Catalog No.S1413 Batch:S141309

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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)
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O

Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.

 5.000mg/ml (8.03mM) 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. 
Clear solution
5% DMSO 95% Corn oil

Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.

 1.650mg/ml (2.65mM) 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. 
* <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, and it is found to inhibit autophagy while inducing apoptosis.
Targets
H+-ATPase [1]
(Cell-free assay)
0.44 nM
In vitro Bafilomycin A1 (Baf-A1) is a toxic macrolide antibiotic derived from Streptomyces griseus, which inhibits the short circuit current induced by the outer mantle epithelium (OME). The IC50 and maximum inhibition dose of this compound are 0.17 μM and 0.5 μM, respectively. [2] In addition, it inhibits the acid influx with an IC50 value of 0.4 nM. It also inhibits the acidification dose-dependently resulting in a lower quenching, and thus a higher fluorescence. [3] This compound prevents the vacuolization of Hela cells induced by H. pylori, with an inhibitory concentration giving 50% of maximal (ID50) of 4 nM, and is very efficient in restoring vacuolated cells to a normal appearance. [4] It also affects the transport of endocytosed material from early to late endocytic compartments, not only dissipating the low endosomal pH but also blocking transport from early to late endosomes in HeLa cells. [5] At doses of 0.1-1 μM, it completely inhibits the acidification of lysosomes revealed by the incubation with acridine orange in BNL CL.2 and A431 cells. [6] When added to Hanks' balanced salt solution, endogenous protein degradation is strongly inhibited and numerous autophagosomes accumulated in H-4-II-E cells. It also prevents the appearance of endocytosed HRP in autophagic vacuoles. [7]
In vivo Bafilomycin A1 (Baf-A1) (1 μM and 0.1 μM) completely inhibits the resorptive activity of cultured osteoclasts. [8] This compound 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 (Baf-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 this compound.
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 Bafilomycin A1 (Baf-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

    --

References

  • https://pubmed.ncbi.nlm.nih.gov/9684329/
  • https://pubmed.ncbi.nlm.nih.gov/15596387/
  • https://pubmed.ncbi.nlm.nih.gov/17576165/
  • https://pubmed.ncbi.nlm.nih.gov/8446034/
  • https://pubmed.ncbi.nlm.nih.gov/9811698/
  • https://pubmed.ncbi.nlm.nih.gov/1832676/
  • https://pubmed.ncbi.nlm.nih.gov/9639028/
  • https://pubmed.ncbi.nlm.nih.gov/7817803/
  • https://pubmed.ncbi.nlm.nih.gov/10574743/
  • https://pubmed.ncbi.nlm.nih.gov/14713959/

Customer Product Validation

Mitochondrial depolarization mediated relocation of mitochondrial RC-TPP into lysosomes and the ensuing lysosomal acidity triggered rhodamine fluorescence. RC-TPP-loaded Tom20-GFP+ HeLa cells were treated without or with CCCP (20 μM) in the presence or absence of BFA (200 nM). Colocalization of Tom20-GFP (in green) and mitochondrial RC-TPP (in blue) is shown in cyan. Scale bar: 10 μm.

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

A549 cells and SK-1 cells co-treated with Baf A1 (100 nM) and SFN-NAC (30 μM) for 24 h, the expression of LC3 I and LC3 II was determined by Western blot.

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

HOS cells were pretreated with Baf-A1 (100 nM) for 2 h prior to treatment with CAP (100 μM) and DDP (16.7 μM) alone or in combination for 24 h. The expression levels of autophagy-related and apoptosis-related proteins were measured by Western blotting (a).

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

Representative immunofluorescence images of TGF-β3-induced MUC5AC in 16HBE cells treated with 3-MA or Baf A1.

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

Selleck's Bafilomycin A1 (Baf-A1) Has Been Cited by 596 Publications

Cytosolic acetyl-coenzyme A is a signalling metabolite to control mitophagy [ Nature, 2025, 10.1038/s41586-025-09745-x] PubMed: 41225001
Oncogenic RAS induces a distinctive form of non-canonical autophagy mediated by the P38-ULK1-PI4KB axis [ Cell Res, 2025, 10.1038/s41422-025-01085-9] PubMed: 40055523
Host FSTL1 defines the impact of stem cell therapy on liver fibrosis by potentiating the early recruitment of inflammatory macrophages [ Signal Transduct Target Ther, 2025, 10(1):81] PubMed: 40050288
LZTR1 regulates epithelial MHC-I expression via NF-κB1 to modulate CD8+ T cells activation [ Cell Discov, 2025, 11(1):84] PubMed: 41162356
Alterations in PD-L1 succinylation shape anti-tumor immune responses in melanoma [ Nat Genet, 2025, 57(3):680-693] PubMed: 40069506
ACSS2 drives senescence-associated secretory phenotype by limiting purine biosynthesis through PAICS acetylation [ Nat Commun, 2025, 16(1):2071] PubMed: 40021646
BiDAC-dependent degradation of plasma membrane proteins by the endolysosomal system [ Nat Commun, 2025, 16(1):4345] PubMed: 40346034
Rab27a regulates the transport of influenza virus membrane proteins to the plasma membrane [ Nat Commun, 2025, 16(1):6271] PubMed: 40623997
The O-glycosyltransferase C1GALT1 promotes EWSR1::FLI1 expression and is a therapeutic target for Ewing sarcoma [ Nat Commun, 2025, 16(1):1267] PubMed: 39894896
S100P is a ferroptosis suppressor to facilitate hepatocellular carcinoma development by rewiring lipid metabolism [ Nat Commun, 2025, 16(1):509] PubMed: 39779666

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SHIPPING AND STORAGE
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