Bafilomycin A1(Baf-A1)

Catalog No.S1413

Bafilomycin A1(Baf-A1) Chemical Structure

Molecular Weight(MW): 622.83

Bafilomycin A1 is a vacuolar H+-ATPase inhibitor with IC50 of 0.44 nM.

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Cited by 28 Publications

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Biological Activity

Description Bafilomycin A1 is a vacuolar H+-ATPase inhibitor with IC50 of 0.44 nM.
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]
Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human MCF7 cells NVr3V3I{TnWwY4Tpc44h[XO|YYm= NYXCS5V[PCCq NIH4TlVKdmirYnn0bY9vKG:oIILhdIFugWOrbj3pcoR2[2WmIHH1eI9xcGGpeTDpckBpfW2jbjDNR2Y4KGOnbHzzJIV5eHKnc4PpcochTUeIUD3MR|Mh[XO|ZYPz[YQh[XNiZHXjdoVie2ViaX6gSWdHWCCuZY\lcJMh[XRiMUCwJI5OKGGodHXyJFQhcHK|IHL5JHdme3Sncn6gZoxwfHSrbnegdoVt[XSrdnWgeI8h[2:wdILvcC=> MnLtNlAxOjhzM{S=
human HeLa cells M2Hke2Z2dmO2aX;uJIF{e2G7 MUC0NFAhdk1? NFrqS4pKdmS3Y4Tpc44hd2ZiYYX0c5Bp[We7IHnuJIh2dWGwIFjlUIEh[2WubIOg[ZhxemW|c3nu[{BGT0[SLVzDN{Bie3Onc4Pl[EBieyCrbnPy[YF{\SCrbjDMR|MuOiCuZY\lcEBifCB2MECgcm0> NFH4VIcyQDN7MUm0PS=>
mouse RAW264.7 cells NHHDN3RCeG:ydH;zbZMh[XO|YYm= MVexNFAhdk1? MlOxNVYhcA>? MUPJcoR2[3Srb36gc4Yh[XCxcITvd4l{KGmwIH3veZNmKFKDV{K2OE44KGOnbHzzJIF{e2W|c3XkJIF{KGyjdHWgZZBweHSxdHnjJINmdGy|IHH0JFExOCCwTTDh[pRmeiBzNjDodpMhfXOrbnegZY5v\XirbjDWMZBzd3CrZHn1cUBqd2SrZHWgd5RicW6rbnegZpkh\myxdzDjfZRwdWW2com= NXPZWXluOTl|MEezOVk>
RAW 264.7 cells MkfkSpVv[3Srb36gZZN{[Xl? MXGxNFAhdk1? NIPLVWtCdnSrbXnjdo9jcWGuIHHjeIl3cXS7IHHnZYlve3RiU3HscY9v\WyuYTDlcpRmemmlYTDUfZBpcW23cnn1cUAyPDB{ODDpcoZm[3SnZDDpckBTSVdiMk[0Mlch[2WubIOgZZN{\XO|ZXSgZZMhcW6lcnXhd4VlKG6rdILpZ{BwgGmmZTDwdo9lfWO2aX;uJIlvKGmwZnXjeIVlKGOnbHzzJIF1KDFyMDDuUS=> NXjLRWRVOTl|MEezOVk>
human H4 cells MoTmSpVv[3Srb36gZZN{[Xl? NWrMPGo1OC52IN88US=> NHjj[I8zPCCq NVHz[GgyUW6mdXP0bY9vKG:oIHzp[4h1KGOqYXnuJFMuT0[SIHzleoVtKGmwIHj1cYFvKEh2IHPlcIx{KGG2IECuOEB2VSCjZoTldkAzPCCqcoOgZpkhcGmpaDD0bJJwfWeqcIX0JIZtfW:{ZYPj[Y5k\SCvaXPyc5Nkd3C7IILlcIF1cX[nIITvJINwdnS{b3y= M1POOFE5ODJ2NUi0

... Click to View More Cell Line Experimental Data
Assay
Methods Test Index PMID
Western blot
Ret / EGFR / p75 NGFR; 

PubMed: 21559479     


PC12 cells were treated with various concentrations of bafilomycin A1 for 24 h. Whole cell lysates (40 µg protein) were subjected to SDS–PAGE and immunoblotted with anti-Ret, anti-p75 NGFR, anti-EGFR, and anti-α-tubulin antibodies.

21559479
Growth inhibition assay
Cell viability; 

PubMed: 20534000     


48 h treatment with bafilomycin A1 (BafA1) decreases cell viability at concentrations ≥ 6 nM . *p<0.05 vs. 0 μM vehicle CTL; #p<0.05 vs. 0-3 nM Baf A1.

20534000
Immunofluorescence
AIF; 

PubMed: 25512644     


Confocal microscopic observation of bafilomycin A1-induced AIF subcellular localization. The nucleus was stained with Hoechst 33258 (blue) and AIF by antibody (red). AIF was observed to relocate from cytoplasmic compartments to the nucleus upon bafilomycin A1 treatment. 

LC3; 

PubMed: 24532803     


PANC1 VC and PANC1 N1 cells were preincubated with control medium or this medium containing the late stage autophagy inhibitor bafilomycin A1 (100 nM) for 30 min at 37℃ followed by a 24-h, 37℃Cincubation with either control medium or this medium containing Dp44mT (5 μM) alone, bafilomycin A1 (100 nM) alone, or the combination of Dp44mT (5 μM) and bafilomycin A1 (100 nM).  Immunofluorescence studies with anti-LC3 antibody. Scale, 20 μm.

25512644 24532803
ELISA
TNF-alpha; 

PubMed: 26240140     


BMDMs were incubated with P. falciparumIRBCs in the presence of bafilomycin A1. TNF-α released into culture medium was analyzed by ELISA.

26240140
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:

[2]
+ Expand

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 Research:

[4]
+ Expand
  • 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.


    (Only for Reference)
Animal Research:

[9]
+ Expand
  • Animal Models: Young (approximately 9 days old) Mozambique tilapia, Oreochromis mossambicus
  • Formulation: Aquarium water containing 0.05 % dimethylsulphoxide (DMSO)
  • Dosages: 10 μM
  • Administration: --
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 6 mg/mL (9.63 mM)
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
Formula

C35H58O9
CAS No. 88899-55-2
Storage powder
in solvent
Synonyms N/A

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Frequently Asked Questions

  • Question 1:

    How to dissolve it？

  • 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.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID