Bafilomycin a1

Manufactured by Abcam

Sourced in United Kingdom, United States

Bafilomycin A1 is a macrolide lactone compound that functions as a specific and potent inhibitor of vacuolar-type H+-ATPase (V-ATPase). V-ATPase is responsible for acidifying various intracellular compartments, including lysosomes, endosomes, and secretory vesicles.

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Lab products found in correlation

Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) 3 methyladenine 3 ma, by Merck Group (2 mentions) Mda mb 231, by American Type Culture Collection (2 mentions) Mda mb 468, by American Type Culture Collection (2 mentions) Rapamycin, by Cell Signaling Technology (1 mentions) 3 methyladenine 3 ma, by Santa Cruz Biotechnology (1 mentions) Chloroquine, by Cell Signaling Technology (1 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions) Hepatocyte growth factor (hgf), by R&D Systems (1 mentions) Hrp conjugated goat anti chicken antibody, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated horse anti mouse igg, by Cell Signaling Technology (1 mentions) Sb366791, by Bio-Techne (1 mentions) Goat anti rabbit igg antibody, by Cell Signaling Technology (1 mentions) Crizotinib, by Bio-Techne (1 mentions) Alexa fluor 488 conjugated donkey anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Hrp conjugated mouse monoclonal antibody against β actin, by Abcam (1 mentions) Rhodamine red x conjugated donkey anti goat igg antibody, by Jackson ImmunoResearch (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Huvecs, by American Type Culture Collection (1 mentions) 3 methyladenine, by Abcam (1 mentions)

Spelling variants of this product

Bafilomycin A1 (ab120497) (6 citations) Bafilomycin A1 (Baf-A1) (4 citations) Bafilomycin (2 citations)

18 protocols using bafilomycin a1

Bovine IFN-γ Modulation Assay

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Bovine IFN-γ was purchased from the Kingfisher Group (King-fisher Biotech, Inc., USA). Bafilomycin A1 and E64d were purchased from Abcam (UK). 3-Methyladenine (3-MA) was purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology Inc., Germany). Chloroquine and rapamycin were purchased from Cell Signaling Technology (USA). The amino acids were purchased from Nanjing Keygen Biotech. Co., Ltd.

Xia X., Che Y., Gao Y., Zhao S., Ao C., Yang H., Liu J., Liu G., Han W., Wang Y, & Lei L. (2016). Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells. Molecules and Cells, 39(5), 410-417.

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Investigating TRPV1 and HGF Signaling

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HGF was from R & D (Minneapolis, MN). 5’-Iodoresiniferatoxin (I-RTX), doxorubicin, crizotinib and SB366791 were from Tocris (R & D, Minneapolis, MN); rabbit monoclonal antibodies to, ERK1/2, CREB, pERK1/2, pCREB, p-Met, CAMKIIαand mouse monoclonal antibodies to c-Met, HRP-conjugated horse anti-mouse IgG and goat anti-rabbit IgG antibody were from Cell Signaling Technology (Danvers, MA); bafilomycin A1, rabbit polyclonal antibodies to TRPV1 and HGF, the goat polyclonal antibody against CGRP, and HRP-conjugated mouse monoclonal antibody against β-actin were from Abcam (Cambridge, UK); Alexa fluor 488-conjugated donkey anti-rabbit IgG and Alexa fluor 680-conjugated donkey anti-goat IgG and rhodamine red-X (RRX)-conjugated donkey anti-goat IgG antibody were from Jackson Laboratory (Bar Harbor, ME); Rhodamine phalloidin was from Life Technologies (Carlsbad, CA); and HRP-conjugated goat anti-chicken antibody was from Santa Cruz Biotechnology (Dallas, TX). Anti-a3V-ATPase antibody was a kind gift from Dr. Sun-Wada, Doshisha Women’s College, Japan.

OKUI T., Hiasa M., Hata K., Roodman G.D., Nakanishi M, & Yoneda T. (2023). The acid-sensing nociceptor TRPV1 controls breast cancer progression in bone via regulating HGF secretion from sensory neurons. Research Square.

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Endothelial Cell Autophagy Regulation

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HUVECs were purchased from ATCC and cultured in RPMI1640 medium, supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) at 37 °C in humidified air containing 5% CO₂. The autophagy inhibitor 3-methyladenine (3-MA, MP, CA, USA) was dissolved in ddH₂O at concentration of 200 mM for storation at room temperature and bafilomycin A1 (Baf A1, Abcam, Cambridge, UK) was dissolved in dimethyl sulfoxide at concentration of 400 mM and kept in 4 °C. The proteasome inhibitor MG132 (Targetmol, MA, USA) was dissolved in dimethyl sulfoxide at stock concentration of 5 mM and kept in −20 °C.The nucleocytoplasmic transport inhibitor leptomycin B (LMB, Beyotime, Shanghai, China) was prepared at concentration of 0.2 mg/mL and stored at −20 °C.The final concentrations of the drugs and the duration of treatments are indicated in the figure legends.

Li C.F., Sun J.X., Gao Y., Shi F., Pan Y.K., Wang Y.C, & Sun X.Q. (2018). Clinorotation-induced autophagy via HDM2-p53-mTOR pathway enhances cell migration in vascular endothelial cells. Cell Death & Disease, 9(2), 147.

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Streptozotocin-Induced Diabetic Rat Model

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Healthy male rats, 6 weeks old, body mass of 180 ∼ 220 g, purchased from Shanghai SLAC, Ltd (Shanghai, China); Streptozotocin (STZ, Sigma, St. Louis, MO, USA); Glucose meter (Roche, Mannheim, Germany); CD68 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); RAW264.7 cells (Shanghai Bogu Biotechnology Co, Shanghai, China); RPMI-1640 medium (Gibco, Waltham, MA, USA); fetal bovine serum (ScienCell, Carlsbad, CA, USA); Dextrose, mannitol, and Rapamycin (RAPA; Sigma, USA); 3-methyladenine (3-MA) (Sigma, USA); Bafilomycin A1 (BAFA; Abcam, Cambridge, UK); Chloroquine (CQ; Sigma, USA); CD68 (Abcam); LC3 (Sigma, USA); Beclin 1 (Abcam); P62 (Cell Signaling Technology, Danvers, MA, USA); Transwell Chambers (Sigma, USA); and Fibronectin (FN; Cloud Clone, Katy, TX, USA).

Jiang Y., Zhao Y., Zhu X., Liu Y., Wu B., Guo Y., Liu B, & Zhang X. (2019). Effects of autophagy on macrophage adhesion and migration in diabetic nephropathy. Renal Failure, 41(1), 682-690.

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Flubendazole induces autophagy-mediated cell death

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Cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). The U937, HL60 and MCF10A cell lines were cultured in Roswell Park Memorial Institute medium (RPMI) 1640 medium and the U87, SW480, RKO, LST174t, HCT116, HepG2, HUH7, SK-HepG1, A549, 786-0, Hela, MCF-7, MDA-MB-231 and MDA-MB-468 were maintained in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (Life Technologies) in 5% CO₂ at 37 °C. Cells were grown to 70-80% confluence in cell culture dishes or plates and all the experiments were performed on logarithmically growing cells.
Flubendazole (SML2510), 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (M2128), 3-MA (M9281), CQ (C6628), DAPI (D9542) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bafilomycin A1 (ab120497) and was purchased from Abcam (Cambridge, UK). Antibodies used in this study were as follow: Beclin1 (3495, CST), SQSTM1/p62 (8025, CST), LC3B (51520, Abcam), Bax (5023, CST), Bcl-2 (2870, CST), Caspase3 (9662, CST), MMP-2 (87809, CST), E-cadherin (14472, CST), HAP1(58600, Abcam), EVE1A (216043, Abcam), HIF1α (36169, CST), β-actin (66009-1-Ig, Proteintech, IL, USA), LAMP1(25630, Abcam), ATG5 (12994, Abcam), Anti-DDDDK tag (Binds to FLAG® tag sequence) (1162, Abcam).

Zhen Y., Zhao R., Wang M., Jiang X., Gao F., Fu L., Zhang L, & Zhou X.L. (2020). Flubendazole elicits anti-cancer effects via targeting EVA1A-modulated autophagy and apoptosis in Triple-negative Breast Cancer. Theranostics, 10(18), 8080-8097.

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Combination Therapy Evaluation

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Cells seeded in 96-well plates (1 × 10⁴ cells/well) were treated with 5 μM USP24-i-101, 24 nM Taxol (paclitaxel, Selleckchem, S1150), 10 nM Baf-A1 (Bafilomycin A1, Abcam, ab120497), and 5 μM CQ (Chloroquine, MedChem Express, HY-17589A) for 24 h. The cell viability was assessed using the PrestoBlue cell viability kit following the manufacturer’s instructions (Thermo Fisher Scientific).

Young M.J., Wang S.A., Chen Y.C., Liu C.Y., Hsu K.C., Tang S.W., Tseng Y.L., Wang Y.C., Lin S.M, & Hung J.J. (2024). USP24-i-101 targeting of USP24 activates autophagy to inhibit drug resistance acquired during cancer therapy. Cell Death and Differentiation, 31(5), 574-591.

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Quantifying Endocytic Pathways in Cells

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Wortmannin, dynasore, SU6655, NH₄Cl, SP600125, IPA-3, MβCD and chlorpromazine were obtained from Sigma (Sigma, MO, United States), Akti-1/2 and bafilomycin A1 were obtained from Abcam (Abcam, Cambridge, United Kingdom), nystatin and EIPA was obtained from Solarbio (Solarbio, Beijing, China).
The rabbit anti-dynamin-2, anti-clathrin heavy chain, anti-Rab5, anti-Rab7, anti-PI3K p85 (phos pho Y458) + PI3 Kinase p55 (phospho Y199), anti-Akt, anti-Src, anti-Src (phospho Y416), anti-phospho-Akt (Ser473) and anti-GAPDH monoclonal antibodies were obtained from Abcam (Abcam, Cambridge, United Kingdom). The rabbit anti-JNK and anti-JNK (Thr183/Tyr185) monoclonal antibodies were obtained from Cell Signaling Technology (Cell Signaling Technology, Danvers, United States). The rabbit anti-PI3 Kinase p85 alpha polyclonal antibody, the mouse anti-Flag monoclonal antibody, goat anti-rabbit and goat anti-mouse HRP-labeled secondary antibody, Alexa fuor-488-conjugated anti-mouse, Cy™3-conjugated anti-rabbit IgG (H + L) and mounting medium with DAPI was purchased from Abcam (Abcam, Cambridge, United Kingdom). The rabbit anti-caveolin-1 monoclonal antibody was obtained from Beyotime (Beyotime Biotechnology, Shanghai, China). The mouse anti-BRSV G protein monoclonal antibody was provided by China Animal Health and Epidemiology Center.

Liu Y., Yang D., Jiang W., Chi T., Kang J., Wang Z, & Wu F. (2024). Cell entry of bovine respiratory syncytial virus through clathrin-mediated endocytosis is regulated by PI3K-Akt and Src-JNK pathways. Frontiers in Microbiology, 15, 1393127.

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Autophagy regulation in breast cancer cells

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MDA-MB-231, MDA-MB-468, and MCF-10A cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). MDA-MB-231 and MCF-10A cells were cultured in DMEM with 10% fetal bovine serum, while MDA-MB-468 cells were cultured in L-15 with 10% fetal bovine serum and incubated with 5% CO₂. Antibodies used in this study were as follows: BRD4 (Cell Signaling Technology, no. 13440S), p-BRD4^S492 (EMD Millipore, ABE no. 1451), c-Myc (Abcam, ab56), LC3B antibody (Abcam, ab51520), SQSTM1/p62 antibody (Abcam, ab56416), CK2α (Cell Signaling Technology, no. 2656S), p-AKT^S129 (Abcam, 133458), GAPDH (Cell Signaling Technology, no. 5174), beclin-1 (Cell Signaling Technology, no. 3495), Bax (Cell Signaling Technology, no. 5023), Bcl-2 (Cell Signaling Technology, no. 2870), caspase-3 (Abcam, ab13847). MTT (M2128), 3-MA (M9281), and Hoechst 33258 were purchased from Solarbio. Bafilomycin A1 (ab120497) was purchased from Abcam.

Kandori H., Kinoshita N., Yamazaki Y., Maeda A., Shichida Y., Needleman R., Lanyi J.K., Bizounok M., Herzfeld J., Raap J, & Lugtenburg J. (2000). Local and distant protein structural changes on photoisomerization of the retinal in bacteriorhodopsin. Proceedings of the National Academy of Sciences of the United States of America, 97(9), 4643-4648.

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Multi-parameter Live Cell Imaging

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Live Cell Imaging solution, Fluo4-AM, CellTracker Deep Red, pHrodo iFL Red STP Ester, NucBlue live cell nuclear stain, NucGreen dead cell nuclear stain, and LIVE/DEAD fixable aqua stain were obtained from Invitrogen. DAPI was from Sigma, and paraformaldehyde (4% in PBS) was obtained from Alfa Aesar. Human annexin V protein was from BD Biosciences, human C5a protein from Peprotech, human TGFβ1 from Miltenyi Biotec, and recombinant human IFNγ and recombinant truncated human vitronectin from Gibco. Cytochalasin D was from Cayman Chemicals, Bafilomycin A1 from Abcam, and Jasplakinolide from Santa Cruz. PSB0739, MRS2179, MRS2211, and cilengitide were all obtained from Tocris. ADP, BSA, and crystal violet were from Sigma.

Hall-Roberts H., Agarwal D., Obst J., Smith T.B., Monzón-Sandoval J., Di Daniel E., Webber C., James W.S., Mead E., Davis J.B, & Cowley S.A. (2020). TREM2 Alzheimer’s variant R47H causes similar transcriptional dysregulation to knockout, yet only subtle functional phenotypes in human iPSC-derived macrophages. Alzheimer's Research & Therapy, 12, 151.

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Modulating Keratinocyte Autophagy Pathways

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The immortalized human keratinocyte cell line HaCaT cell from the Institute of Cell (Shanghai, China) was cultured in DMEM (Gibco, America) supplemented with 10% fetal calf serum (Life Technologies, America), 1% penicillin and 1% streptomycin at standard conditions (37°C, 5% CO₂, in a humidified atmosphere). Additionally, Normal Human Epidermal Keratinocytes (NHEKs), purchased from PromoCell (Heidelberg, Germany, C-12003), were cultured in the company’s supporting medium. 1×10⁵ cells/well were seeded onto six-well plates. The cells were left untreated or treated with 50nM Rapamycin (Abcam, UK) for both HaCaT cells and NHEKs, or 100nM Bafilomycin A1 (Abcam, UK) for HaCaT cells and 50nM for NHEKs. The cells were then collected according to the indicated period.

Geng Q., Wei G., Hu Y., Xu J, & Song X. (2023). Alterations of Autophagy Modify Lipids in Epidermal Keratinocytes. Clinical, Cosmetic and Investigational Dermatology, 16, 1569-1581.

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