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Bafilomycin A1

Bafilomycin A1 is a macrolide antibiotic that acts as a potent and specific inhibitor of vacuolar-type H+-ATPases.
It is widely used in cell biology research to study the role of acidic compartments in cellular processes.
Bafilomycin A1 has been shown to disrupt lysosomal and endosomal function, leading to changes in cellular signaling, trafficking, and metabolism.
Resaerchers can use PubCompare.ai's AI-powered optimization to easily locate high-quality, reproducible protocols for working with Bafilomycin A1 from the scientific literature, preprints, and patents.
This can help ensure more reliable and efficeint research outcomes when investigating the cellular effects of this important pharmacological tool.

Most cited protocols related to «Bafilomycin A1»

1
Comprehensive Resource of Natural Compounds
Cited 20 times
A chemical library of 658-natural compounds was kindly provided by Dr. Sang Jeon Chung of Sungkyunkwan University (Suwon, Korea). Kaempferide (69545), dimethylsulfoxide (D2650), bafilomycin A1 (B1793), rapamycin (553210), tiliroside (79257), chloroquine (C6628), orlistat (O4139), palmitic acid (P5585), oleic acid (O1383), acridine orange (A6014), oil-red-O (O0625), dexamethasone (D8893), insulin (I0516), and 3-isobutyl-1-methylxanthine (I5879) were purchased from Sigma-Aldrich. BODIPY 493/503 (D3922), Hoechst33342 (H3570), lipofectamine LTX (94756), lipofectamine 2000 (52887), Plus reagent (10964), protease and phosphatase inhibitor solution (78441), M-PER kit (89842Y), DMEM, fetal bovine serum (FBS), bovine serum, and antibiotics were purchased from Invitrogen ThermoFisher Scientific. For in vivo experiments, Kaempferide (K0057) was purchased from TCI Chemicals. siRNA targeting TUFM was purchased from Dharmacon. mRFP-GFP-LC3B plasmids were kindly provided by Dr. Jaewhan Song of Yonsei University (Seoul, Korea).
Kim D., Hwang H.Y., Ji E.S., Kim J.Y., Yoo J.S, & Kwon H.J. (2021). Activation of mitochondrial TUFM ameliorates metabolic dysregulation through coordinating autophagy induction. Communications Biology, 4, 1.
Publication 2021
1-Methyl-3-isobutylxanthine 4,4-difluoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene Acridine Orange Antibiotics, Antitubercular bafilomycin A1 Bos taurus Chloroquine Dexamethasone Fetal Bovine Serum Hoechst33342 Insulin kaempferide Lipofectamine lipofectamine 2000 Oleic Acid Orlistat Palmitic Acid Peptide Hydrolases Phosphoric Monoester Hydrolases Plasmids RNA, Small Interfering Serum Sirolimus solvent red 27 Sulfoxide, Dimethyl tiliroside
2
Fluorescence Imaging of Lysosomal and Autophagic Markers
Cited 19 times
For acridine orange (AO) stain in the screen, HeLa cells were grown in 96-well plates. Cells were treated with DMSO or indatraline, bafilomycin A1, and 658-natural chemicals for 24 h, and stained with 5 μg/mL AO. Fluorescence intensity was measured by victor plate reader, where fluorescence intensity of each well in the plate is promptly displayed as numerical readout. For confocal microscopy, 3T3-L1 or HeLa cells were grown on 15 mm coverslips at a density of 1.0 × 105 cells/well in 6-well plates The cells were then treated with drugs for the time periods indicated, followed by treatment with 5 μg/mL AO (Sigma-Aldrich). Nuclei were stained with Hoechst. Following incubation for 20 min, the cells were fixed with 4% PFA and washed three times with PBS. Images were obtained using an LSM880 confocal microscope at ×400 magnification. Red fluorescence intensity was quantified using Image J2 software. For BODIPY FL-Pepstatin A stain, HeLa cells were grown on 15 mm coverslips at a density of 1.0 × 105 cells/well in 6-well plates. The cells were then treated with drugs for the time periods indicated, followed by treatment with 1 μM BODIPY FL-Pepstatin A (Invitrogen) for 30 min. Nuclei were stained with Hoechst. Following incubation for 20 min, the cells were fixed with 4% PFA and washed three times with PBS. Images were obtained using an LSM880 confocal microscope at 400× magnification. For DQ-BSA analysis, HeLa cells were grown on 15 mm coverslips at a density of 1.0 × 105 cells/well in 6-well plates. The cells were then treated with 10 μg/mL DQ-BSA for 2 h. After change medium, cells were treated with drugs for the time periods indicated. Nuclei were stained with Hoechst with incubation for 20 min, the cells were fixed with 4% PFA and washed three times with PBS. Images were obtained using an LSM880 confocal microscope at ×400 magnification.
Kim D., Hwang H.Y., Ji E.S., Kim J.Y., Yoo J.S, & Kwon H.J. (2021). Activation of mitochondrial TUFM ameliorates metabolic dysregulation through coordinating autophagy induction. Communications Biology, 4, 1.
Publication 2021
3T3-L1 Cells Acridine Orange bafilomycin A1 BODIPY Cell Nucleus Cells Fluorescence HeLa Cells indatraline Microscopy, Confocal pepstatin Pharmaceutical Preparations Sulfoxide, Dimethyl
3
Mitophagy Assay using mito-QC Reporter
Cited 19 times
Experiments were conducted in either SH-SY5Y or ARPE-19 cells following American Type Culture collection (ATCC) guidance. In brief, cells were cultured in 1:1 DMEM:F-12 media (Thermo Scientific; #12634010) supplemented with 10 % (v/v) FBS (Sigma; #F7524), 2 mM l-glutamine (Thermo Scientific; #25030-024), 100 U/ml penicillin and 0.1 mg/ml streptomycin (Thermo Scientific; #15140-122) and incubated at 37 °C with 5 % CO2 in a water-saturated incubator. The mito-QC reporter was retrovirally transfected as previously described (Allen et al., 2013 (link)). Mitophagy was assessed from pools of cells transfected with the reporter. Iron chelation induced mitophagy with Deferiprone (DFP) (Sigma; #379409) was achieved as previously reported, where cells were treated with 1 mM DFP, dissolved in water before use with a pulse of heat (90 °C), for 24 h. For hypoxia treatment cells were transferred to a Ruskinn INVIVO2 300 workstation (Ruskinn/Baker) at 37 °C in 1 % O2 and 5 % CO2, where were incubated for 42 h. After 36 h incubation in hypoxia, 50 nM of Bafilomycin A1 (Enzo; BML-CM110) was added to inhibit lysosome acidification during the remaining 6 h up to 42 h.
The experiments using the mito-QC reporter to assess mitophagy were processed as previously described (Allen et al., 2013 (link)). In brief, cells containing the mito-QC reporter were washed once with PBS (Thermo Scientific; #14190-094) and fixed with 3.7 % PFA (Sigma; P6148) in 200 mM HEPES (Formedium; HEPES10), pH 7.0 for 10 min at room temperature. Then, samples were washed twice with DMEM media supplemented with 10 mM HEPES pH 7.0 and 0.04 % Sodium Azide (Sigma; S8032), followed by a 10 min incubation. After two washes with PBS, coverslips were incubated with 1 μg/ml Hoechst (Thermo Scientific; #62249) for 30 min, washed three times with PBS, dipped in MilliQ water and mounted onto glass slides with ProLong Diamond Antifade Mountant (Thermo Scientific; #P36961).
The immunostaining of SH-SY5Y mito-QC reporter cells with LAMP1 was performed as follow: cells were washing once with PBS and fixing cells with 3.7 % PFA pH 7.0 for 20 min at room temperature. The cross-linking reaction was quenched as described above. Cells were the permeabilised with 1 % BSA (Roche; #10735108001)/PBS + 0.1 % NP-40 (Merck/Millipore; #492016) for 3 min at room temperature. In contrast to tissue preparations, the permeabilization of cell in vitro can affect the red-only mitolysosome stability. Therefore, the quantification of mitophagy using the mito-QC reporter should be avoided after detergent permeabilization of cells in vitro. Following permeabilization and fixation, coverslips were next washed two times with 1 % BSA/PBS and followed by a 30 min incubation at room temperature. The LAMP1 primary antibody (Santa Cruz; sc-20011) were incubated at 1/300 in 1 % BSA/PBS for 1.5 h at 37 °C. Then, coverslips were washed with 1 % BSA/PBS four times of 10 min each. The secondary anti-mouse Alexa Fluor 647 nm antibody (Thermo Scientific; #A21236) was incubated at 1/1000 in 1 % BSA/PBS for 30 min in the dark at room temperature. Coverslips were washed again four times with 1 % BSA/PBS for 10 min each. Cells were then stained with Hoechst and mounted glass slides as described above.
Images with the mito-QC reporter to quantify mitophagy were acquired using a Nikon Eclipse Ti widefield microscope (Plan Apo Lambda 60x Oil Ph3 DM) with the NIS-Elements software, while the images for the LAMP1 co-localisation experiment were acquired in a Leica SP8 laser scanning confocal microscope (HC PL APO 63x/1.40 oil CS2). All the images were processed with FIJI v1.52n software (ImageJ, NIH). Quantification of mitophagy was performed from three independent experiments counting over 50 cells for condition. Images were processed with the mito-QC Counter. For images acquired in the widefield microscope the following parameters were used: Radius for smoothing images = 1, Ratio threshold = 0.6, and Red channel threshold = mean + 0 standard deviation. For images acquired in the Leica SP8 confocal microscope the following parameters were used: Radius for smoothing images = 2, Ratio threshold = 1.5, and Red channel threshold = mean + 0.5 standard deviation.
Montava-Garriga L., Singh F., Ball G, & Ganley I.G. (2020). Semi-automated quantitation of mitophagy in cells and tissues. Mechanisms of Ageing and Development, 185, 111196.
Publication 2020
4
Comprehensive Bladder Cancer Research Protocol
Cited 8 times

Plasmids, Reagents, and Antibodies: The plasmid of the human ATG7 promoter (from −1398 to −227)‐driven luciferase reporter was constructed with KpnI and BglII using genomic DNA purified from UMUC3 cells (Figure S5, Supporting Information). Human ATG7 3′‐UTR luciferase reporter has already been described (Figure S6A, Supporting Information).39 ATG7 3′‐UTR mutant luciferase reporter (MIR190A binding site was mutated) was cloned into the pMIR luciferase reporter. V5‐DEST‐MIR190A and its control construct were kindly gifted by Dr. Ping‐Yee Law (Department of Pharmacology, University of Minnesota, Minneapolis, MN, USA). The plasmid of ARHGDIB promoter‐driven luciferase reporter was synthesized by Cyagen Bioscience and was inserted into pGL3 Basic plasmid. The GFP‐tagged ARHGDIB was kindly gifted by Dr. Martin A. Schwartz (Robert M. Berne Cardiovascular Research Center, University of Virginia, Charlottesville, Virginia).42 The Myc‐ATG7 construct was obtained from Addgene (Cambridge, MA). The GFP‐ATG7 plasmid was constructed with KpnI and NotI using cDNA purified from UMUC3 cells. MIR196B constitutively expressed plasmid was kindly gifted from Dr. Wen‐Chang Lin (Institute of Biomedical Sciences, Academic Sinica, Nankang, Taipei 115, Taiwan, Republic of China).43 The HA‐tagged HNRNPD (38066) and ATG7 constitutively expressed plasmid (24921) were obtained from Addgene (Cambridge, MA, USA). The plasmids of short hairpin RNA specifically targeting ATG7 (shATG7) (human, RHS4531‐EG10533), HNRNPD (shHNRNPD) (human, RHS4531‐EG3184), and the plasmid of antisense of MIR190A (HmiR‐AN0258‐AM03) were purchased from GeneCopoeia (Rockville, MD, USA). All plasmids were prepared by the Plasmid Preparation/Extraction Maxi Kit (10063) from Qiagen (Valencia, CA, USA). BBN (B0938) was purchased from TCI AMERICAN (Cambridge, MA, USA). The chemicals Cycloheximide (CHX) (239763) and Act D (129935) were purchased from Calbiochem (Billerica, MA, USA). Bafilomycin A1 (sc‐201550A) was bought from Santa Cruz (St. Louis, MO, USA). The antibodies of GAPDH (5174S), RHOA (2117S), RAC1,2,3 (2465S), GFP (2956S), ELAVL1 (12582S), and Autophagy Antibody Sample Kit (4445S) were purchased from Cell Signaling Technology (Beverly, MA, USA). The antibody specific against HNRNPD (OAAF01114) was bought from Aviva Systems Biology (San Diego, CA, USA). Antibodies against NCL (sc‐13057), ARHGDIA (sc‐6047), and ARHGDIB (sc‐32227) were bought from Santa Cruz (Dallas, Texas 75220 USA), while antibodies against β‐Actin (A5316) were purchased from Sigma (St. Louis, MO, USA).
Cell Lines and Cell Culture: UROtsa, UMUC3, and T24 cells were used in the previous studies.44 Before/after utilization for research, PowerPlex 16 HS System was used to do DNA tests and authenticate all the cell lines by Genetica DNA Laboratories (Burlington, NC, USA).
Western Blot Analysis: The whole BC cell extracts were prepared and treated as described in the previous studies.45 The images were acquired by scanning with the phosphor imager (Typhoon FLA 7000, GE, Pittsburgh, PA).
Transfection and Luciferase Assay: In Vitro Transfection Reagent PolyJet DNA (SL100468) (SignaGen Laboratories, Rockville, MD, USA) was used and described in the previous studies.45 The luciferase reporters and pRL‐TK were transiently transfected into the indicated cells. 24 h after transfection, the luciferase Assay System kit (E1960) (Promega, Madison, WI, USA) was used to determine the luciferase activity. The results were normalized by internal TK signal. All experiments were done in triplicate and the results expressed as mean ± standard error.
Reverse Transcription‐Polymerase Chain Reaction (RT‐PCR): Total RNA was extracted using the TRIzol reagent (15596026) (Invitrogen, Grand Island, NY, USA), and then. 5.0 µg RNA was used for first‐strand cDNA synthesis with oligo (dT) 20 primer by using Super‐Script First‐Strand Synthesis system (18080051) (Invitrogen, Grand Island, NY, USA). The PCR product was analyzed by agarose gel. The densitometric analyses of the product bands were performed using the Image Quant 5.2 software (GE Healthcare, Pittsburgh, PA, USA). The primers used in this study were: human ATG7 (Forward, 5′‐GCC AAG ATC TCC TAC TCC AATC‐3′; Reverse, 5′‐CAG AAG TAG CAG CCA AGC TTGT‐3′) and human β‐Actin (Forward, 5′‐CTC CAT CCT GGC CTC GCT GT‐3′; Reverse, 5′‐GCT GTC ACC TTC ACC GTT CC‐3′).
Quantitative RT‐PCR for mRNA and miRNA Assay: Fast SYBR Green Master Mix kit (Applied Biosystems, 4385614) was used to conduct real‐time PCR in the 7900HT Fast Real‐Time PCR System (Applied Biosystems, Foster City, CA, USA). The primers used were: human ARHGDIB (Forward, 5′‐ACC CGG CTC ACC CTG GTT TGT‐3′; Reverse, 5′‐ACC CCA GTC CTG TAG GTG TGC TG‐3′); human HNRNPD (Forward, 5′‐AAA TTG AAT GGG AAG GTG AT‐3′; Reverse, 5′‐GAA CCC ACG CCT CTT ATT G‐3′), and human β‐Actin (Forward, 5′‐CTC CAT CCT GGC CTC GCT GT‐3′; Reverse, 5′‐GCT GTC ACC TTC ACC GTT CC‐3′). The primer for MIR190A (MS00011333) was purchased from Qiagen (Germantown, MD, USA).
RNA Immunoprecipitation (RNA‐IP) Assay: 293T cells were transiently transfected with HA‐HNRNPD. Then, the RNA‐IP assay was performed as described in the studies previously.46 RT‐PCR was performed to identify the mRNA presented in the immune‐complex.
Cell Migration and Invasion Assay: Control inserts without matrigel and permeable support for 24‐well plate with 8.0 µm transparent PET membrane were purchased from Corning Incorporated (Corning, NY, USA) (353097), and the invasion kit (354480) was purchased from BD Biosciences (Bedford, MA, USA). The cell migration and invasion activity of the indicated cells were evaluated as described in the previous studies.47 The images of the results were taken under a microscopy, Olympus DP71 (Olympus America Inc. Center Valley, PA, USA), and the number of the cells in each image was counted by the software “Image J.” The invasion rate was normalized with the insert control according to the manufacturer's instruction.
The Construct of Human ATG7 Promoter‐Driven Luciferase Reporter, ATG7 mRNA 3′‐UTR Mutant Luciferase Reporter, and pLentiIII‐GFP‐ATG7 Plasmid: The plasmid of ATG7 promoter (from −1398 to −227)‐driven luciferase reporter was constructed by amplifying from genomic DNA isolated from UMUC3 cells based on the NCBI database, using primers: forward, 5′‐ACT GGT ACC ACT GAC ACA CAC AAC CCC CTA CTG AG‐3′ and Reverse, 5′‐ACT AGA TCT GAG AGG CGG CAT CAA ACG CAG CAC A‐3′, and then subcloned into the KpnI and BglII sites of the PGL3‐basic vector, thus originating the ATG7 promoter‐driven luciferase reporter plasmid. The relevant sequence of human ATG7 promoter (from −1398 to −227) was indicated in Figure S5 in the Supporting Information. The seed region of putative MIR190A/ATG7 interacting sequence (see Figure 2P) was introduced with a three‐point mutation to produce the pMIR‐ATG7 3′‐UTR mutant plasmid by using primers: MIRMUTFOR, 5′‐AAT TAA ATA GCT ACA GCG CAT TAA CAA ATT AAT GTT C‐3′ and MIRMUTREV, 5′‐GAA CAT TAA TTT GTT AAT GCG CTG TAG CTA TTT AAT T ‐3′. The plasmid of pLentiIII‐GFP‐ATG7 was constructed by amplifying from cDNA isolated from UMUC3 cells, using primers: forward, 5′‐ CGG GGT ACC CAA GAA ATA ATG GCG GCA G‐3′ and Reverse, 5′‐ ATA AGA ATG CGG CCG CAT CTC AGA TGG TCT CAT C‐3′, and then subcloned into the KpnI and NotI sites of the pLentiIII‐GFP vector, thus originating the pLentiIII‐GFP‐ATG7 plasmid. Constructs were all sequence verified by GENEWIZ (South Plainfield, NJ, USA).
Human Bladder Cancer Tissue Specimens: All human studies were performed in compliance with the relevant laws and institutional guidelines, and were approved by the Ethics Committee of Wenzhou Medical University (March 6, 2016). Informed consent was obtained from all patients before sample collection. All human bladder cancer tissue specimens were obtained from patients who underwent radical cystectomy at the Department of Urology of the Union Hospital of Tongji Medical College (Wuhan, China) during 2012 and 2013 and the First Affiliated Hospital of Wenzhou Medical University (Wenzhou, China) between 2015 and 2016, which have been described in the previous study.48Tumor Xenografts and In vivo BBN Treatment of Mice: In accordance with NIH guidelines, all animal procedures were approved by the New York University Committee on Animal Resources. The tumor xenograft studies were completed in the Animal Institute of Wenzhou Medical University, which is approved by the Medical Experimental Animal Care Commission of Wenzhou Medical University. The experiment of mice treated with BBN in vivo and the tumor xenograft studies were described in the previous study.44 None of the mice were sacrificed or died before the end of the experiment.
Immunohistochemistry Paraffin (IHC‐P) of Mouse and Human Bladder Specimens: Mouse or human bladder cancer specimens were formalin‐fixed and paraffin‐embedded. For IHC staining, antibodies specific were used against ATG7 (sc‐33211) (Santa Cruz St. Louis, MO, USA), NCL (sc‐17826) (Santa Cruz St. Louis, MO, USA), HNRNPD (OAAF01114) (Aviva Systems Biology, San Diego, USA), or ARHGDIB (sc‐32227) (Santa Cruz St. Louis, MO, USA). The AxioVision Rel.4.6 computerized image analysis system (Carl Zeiss, Oberkochen, Germany) was used to get the resultant immunostaining images. Image‐Pro Plus version 6.0 (Media Cybernetics, MD, USA) was used to analyze the protein expression by calculating the integrated optical density per stained area (IOD/area).
Statistical Methods: Student's t‐test was utilized to determine the significance of differences between different groups. The differences were considered to be significant at p < 0.05.
Zhu J., Tian Z., Li Y., Hua X., Zhang D., Li J., Jin H., Xu J., Chen W., Niu B., Wu X., Comincini S., Huang H, & Huang C. (2019). ATG7 Promotes Bladder Cancer Invasion via Autophagy‐Mediated Increased ARHGDIB mRNA Stability. Advanced Science, 6(8), 1801927.
Publication 2019
5'-chloroacetamido-5'-deoxythymidine Actins Anabolism Animals Animals, Laboratory Antibodies Autophagy bafilomycin A1 Binding Sites Biological Assay Cancer of Bladder Cardiovascular System Cell Culture Techniques Cell Extracts Cell Lines Cells Cloning Vectors Complex, Immune Cycloheximide Cystectomy Dactinomycin Densitometry DNA, Complementary Ethics Committees Fast Green Formalin GAPDH protein, human Genome HEK293 Cells Homo sapiens Immunoglobulins Immunohistochemistry Immunoprecipitation Luciferases matrigel MicroRNAs Microscopy Migration, Cell Mus Neoplasms Oligonucleotide Primers Oligonucleotides Paraffin Paragangliomas 3 Patients Permeability Phosphorus Plasmids Point Mutation Promega Proteins Reverse Transcriptase Polymerase Chain Reaction RHOA protein, human RNA, Messenger Sepharose Short Hairpin RNA Specimen Collection Tissue, Membrane Tissues Transfection trizol Typhoons Urinary Bladder Vision Western Blot Xenografting
5
Cardiomyocyte Hypoxia-Reoxygenation Model
Cited 7 times
The HR injury model was mimicked in vitro by 45 min of hypoxia and 6 h of reoxygenation. The primary cardiomyocytes were isolated from WT and CK2αCKO mice according to our previous study [44 (link)]. To inhibit the mitophagy, siRNA against FUNDC1 was transfected to cardiomyocytes isolated from WT mice. To observe the autophagic flux, Bafilomycin-A1 (0.5 μM, Selleck Chemicals) was used 12 h before treatment. Details on MTT assay, TUNEL staining and caspase3/9 activities are described in the supplemental methods. The siRNAs specific against the expression of CK2α and FUNDC1 or control siRNAs were purchased Santa Cruz Biotechnology. The siRNA transfection was based on our previous study [45 (link)], and the transfection efficiency was confirmed by western blots.
Zhou H., Zhu P., Wang J., Zhu H., Ren J, & Chen Y. (2018). Pathogenesis of cardiac ischemia reperfusion injury is associated with CK2α-disturbed mitochondrial homeostasis via suppression of FUNDC1-related mitophagy. Cell Death and Differentiation, 25(6), 1080-1093.
Publication 2018
Autophagy bafilomycin A1 Biological Assay Cardiac Arrest Caspase 3 CSNK2A1 protein, human Hypoxia Injuries In Situ Nick-End Labeling Mitophagy Mus Myocytes, Cardiac RNA, Small Interfering Transfection Western Blot

Most recents protocols related to «Bafilomycin A1»

1
Morphological Analysis of Autophagy and DGAT1 Inhibitors
In experiments employing autophagy inhibitors, embryos were treated in embryo medium from 72 to 120 h post-fertilization (hpf) for a morphological analysis, with Bufilomycin A1 (2.5 nM; EMD Millipore, Darmstadt, Germany) or dimethyl sulfoxide (DMSO) as a control. For experiments utilizing DGAT1 inhibitors, embryos were similarly treated in embryo medium from 72 to 120 hpf for the morphological analysis, with A922500 (2 mM; Sigma-Aldrich, St. Louis, MO, USA) or DMSO as a control. The medium containing the compounds was changed daily.
Inoue M., Sebastian W.A., Sonoda S., Miyahara H., Shimizu N., Shiraishi H., Maeda M., Yanagi K., Kaname T., Hanada R., Hanada T, & Ihara K. (2024). Biallelic variants in LARS1 induce steatosis in developing zebrafish liver via enhanced autophagy. Orphanet Journal of Rare Diseases, 19, 219.
Publication 2024
2
Autophagic Flux Analysis in C2C12 Cells
Autophagic flux was determined by the measure of LC3 turnover by treating cells with bafilomycin A1 (11038, Cayman, MI, USA), which blocks the fusion of autophagosomes with lysosomes inhibiting the late degradation stage of autophagy. Early- and late-passage C2C12 control cells were seeded at a density of 5 × 103 cells/cm2 and treated 24 h later with 0.2 μM bafilomycin A1 during 24 h. After 4- and 7-days differentiation, early- and late-passage Dif4d and Dif7d cells were treated with 0.2 μM bafilomycin A1 during 24 h. LC3 and SQSTM1/p62 were detected by western blot analysis, as described above.
Potes Y., Bermejo-Millo J.C., Mendes C., Castelão-Baptista J.P., Díaz-Luis A., Pérez-Martínez Z., Solano J.J., Sardão V.A., Oliveira P.J., Caballero B., Coto-Montes A, & Vega-Naredo I. (2024). p66Shc signaling and autophagy impact on C2C12 myoblast differentiation during senescence. Cell Death & Disease, 15(3), 200.
Publication 2024
3
Autophagy Inhibitors: Preparation and Application
Autophagy inhibitors were prepared in stock concentration according to the manufacturer’s instructions: chloroquine diphosphate (Sigma-Aldrich, C6628), wortmannin (Sigma-Aldrich, W1628), 3-methyladenine (3MA, Cayman, 13242), and bafilomycin A1 (Cayman, 11038). The inhibitors were then diluted into each working concentration with cell culture medium and applied into media reservoir a day or two days after EC loading. Two types of control medium were set considering the addition of DMSO as solvent for wortmannin, 3MA, and bafilomycin A1; control medium (EGM-2) and 0.5% DMSO control medium.
Lee S., Kim H., Kim B.S., Chae S., Jung S., Lee J.S., Yu J., Son K., Chung M., Kim J.K., Hwang D., Baek S.H, & Jeon N.L. (2024). Angiogenesis-on-a-chip coupled with single-cell RNA sequencing reveals spatially differential activations of autophagy along angiogenic sprouts. Nature Communications, 15, 230.
Publication 2024
4
Pretreatment Effects on OGD/R Injury
Chloroquine (CQ) and bafilomycin A1 (Baf A1) were purchased from Selleck Chemicals (Houston, TX, USA). MG132 was purchased from Sigma-Aldrich. HCMECs were pretreated with MG132 (10 μM), CQ (20 μM), and Baf A1 (500 ng/mL) for 2 h before OGD/R injury treatment.
Liu M., Li S., Yin M., Li Y., Chen J., Chen Y., Zhou Y., Li Q., Xu F., Dai C., Xia Y., Chen A., Lu D., Chen Z., Qian J, & Ge J. (2024). Pinacidil ameliorates cardiac microvascular ischemia–reperfusion injury by inhibiting chaperone-mediated autophagy of calreticulin. Basic Research in Cardiology, 119(1), 113-131.
Publication 2024
5
V-ATPase Modulates Drug Resistance
To determine V-ATPase’s role in drug resistance, A549 cells were treated with 20 μM sunitinib (a target drug) + 0.5 mM ATP and/or Bafilomycin A1 (100, 200 nM), an inhibitor of V-ATPase (37 (link), 38 (link)), and incubated for 24 hours; cell viability/proliferation was measured by resazurin assay. After the contribution of V-ATPase in the drug resistance was shown, A549 cells were treated with 0.5 mM ATP and/or Bafilomycin A1 (100, 200 nM) and incubated for 2, 4, 6, and 8 hours and iATP levels were measured by an ATP uptake assay to assess V-ATPase’s role in elevating iATP levels.
Zhang H., Song J., Ward R., Han Y., Hunt A., Shriwas P., Steed A., Edwards C., Cao Y., Co M, & Chen X. (2024). Diverse temporal and spatial mechanisms work, partially through Stanniocalcin-1, V-ATPase and senescence, to activate the extracellular ATP-mediated drug resistance in human cancer cells. Frontiers in Oncology, 14, 1276092.
Publication 2024

Top products related to «Bafilomycin A1»

1
Bafilomycin a1 by Merck Group
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Bafilomycin A1 is a macrolide compound that acts as a potent and specific inhibitor of vacuolar-type H+-ATPases (V-ATPases). V-ATPases are involved in the acidification of various intracellular compartments, making Bafilomycin A1 a useful tool for studying cellular processes that rely on pH regulation.
2
Chloroquine by Merck Group
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Chloroquine is a laboratory chemical primarily used as a research tool in biochemical and cell biology applications. It is a white, crystalline solid that is soluble in water. Chloroquine is commonly used in experiments to study cellular processes, such as autophagy and endocytosis, by inhibiting the function of lysosomes. Its core function is to serve as a research reagent for scientific investigations, without making any claims about its intended use.
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Rapamycin is a macrolide compound isolated from the bacterium Streptomyces hygroscopicus. It functions as an immunosuppressant and has anti-proliferative effects.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Cycloheximide (chx) by Merck Group
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Dimethyl sulfoxide (dmso) by Merck Group
Sourced in United States, Germany, United Kingdom, China, Italy, Sao Tome and Principe, France, Macao, India, Canada, Switzerland, Japan, Australia, Spain, Poland, Belgium, Brazil, Czechia, Portugal, Austria, Denmark, Israel, Sweden, Ireland, Hungary, Mexico, Netherlands, Singapore, Indonesia, Slovakia, Cameroon, Norway, Thailand, Chile, Finland, Malaysia, Latvia, New Zealand, Hong Kong, Pakistan, Uruguay, Bangladesh
DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
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Bafilomycin a1 by Selleck Chemicals
Sourced in United States, China
Bafilomycin A1 is a macrolide lactone compound that acts as a specific inhibitor of vacuolar-type H+-ATPase (V-ATPase). It is commonly used as a research tool in cell biology and biochemistry studies.
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Dulbecco modified eagle medium (dmem) by Thermo Fisher Scientific
Sourced in United States, China, United Kingdom, Germany, France, Australia, Canada, Japan, Italy, Switzerland, Belgium, Austria, Spain, Israel, New Zealand, Ireland, Denmark, India, Poland, Sweden, Argentina, Netherlands, Brazil, Macao, Singapore, Sao Tome and Principe, Cameroon, Hong Kong, Portugal, Morocco, Hungary, Finland, Puerto Rico, Holy See (Vatican City State), Gabon, Bulgaria, Norway, Jamaica
DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.
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Lipofectamine 2000 by Thermo Fisher Scientific
Sourced in United States, China, Germany, United Kingdom, Canada, Japan, France, Italy, Switzerland, Australia, Spain, Belgium, Denmark, Singapore, India, Netherlands, Sweden, New Zealand, Portugal, Poland, Israel, Lithuania, Hong Kong, Argentina, Ireland, Austria, Czechia, Cameroon, Taiwan, Province of China, Morocco
Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.

More about "Bafilomycin A1"

Bafilomycin A1 is a powerful and specific inhibitor of vacuolar-type H+-ATPases, a class of proton pumps that play a crucial role in maintaining the acidic environment of cellular compartments like lysosomes and endosomes.
This macrolide antibiotic is widely utilized in cell biology research to investigate the functional significance of these acidic organelles in various cellular processes, such as signaling, trafficking, and metabolism.
Researchers often employ Bafilomycin A1 in combination with other pharmacological agents like Chloroquine, Rapamycin, MG132, and Cycloheximide to dissect the complex interplay between cellular pathways and organelle function.
By disrupting lysosomal and endosomal acidification, Bafilomycin A1 can lead to significant changes in cellular homeostasis, protein degradation, and autophagy, making it a valuable tool for understanding the role of these fundamental cellular mechanisms.
To ensure reliable and efficient research outcomes when working with Bafilomycin A1, scientists can utilize PubCompare.ai's AI-powered optimization to easily locate high-quality, reproducible protocols from the scientific literature, preprints, and patents.
This platform allows researchers to compare and identify the most accurate and robust procedures for working with Bafilomycin A1, as well as other related compounds like DMSO and DMEM, which are often used in cell culture experiments.
By leveraging PubCompare.ai's advanced capabilities, researchers can experience more reliable and efficient investigations into the cellular effects of Bafilomycin A1 and other important pharmacological tools, ultimately contributing to a deeper understanding of fundamental biological processes and their implications in health and disease.