Lysotracker red dnd 99 l 7528

Manufactured by Thermo Fisher Scientific

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LysoTracker Red DND-99 (L-7528) is a fluorescent dye that selectively stains acidic organelles, such as lysosomes, in live cells. It is a useful tool for visualizing and studying lysosomal dynamics in biological research.

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Lab products found in correlation

Hrp conjugated goat anti rabbit igg h l, by Bio-Rad (2 mentions) Fluorescein conjugated dextran, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Lc3b l7543, by Merck Group (1 mentions) Flag f7425, by Merck Group (1 mentions) Ab27156, by Abcam (1 mentions) N acetyl l cysteine, by Merck Group (1 mentions) Gfp g10362, by Thermo Fisher Scientific (1 mentions) Bafilomycin a1, by Fujifilm (1 mentions) Dulbecco s modified eagle s medium dmem 11 965 118, by Thermo Fisher Scientific (1 mentions) Fluorescence labeled secondary antibody, by Jackson ImmunoResearch (1 mentions) Fetal bovine serum fbs 16 000 044, by Thermo Fisher Scientific (1 mentions) Gapdh sc 25778, by Santa Cruz Biotechnology (1 mentions) Dapi h 1200, by Vector Laboratories (1 mentions) Resazurin, by Merck Group (1 mentions) Fk506, by Merck Group (1 mentions) 4 pba, by Merck Group (1 mentions) Ceapin a7, by Merck Group (1 mentions) Anti gm130 610822, by BD (1 mentions) Anti xbp1 sc 7160, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

LysoTracker Red DND-99 (961 citations) LysoTracker Red (467 citations) LysoTracker (310 citations) LysoTracker DND-99 (39 citations) LysoTracker Red DND (11 citations) Red DND-99 (4 citations) Lysotracker DND-99 (L-7528) (2 citations)

9 protocols using lysotracker red dnd 99 l 7528

Evaluating Mitochondrial Function and Oxidative Stress

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Puromycin (P8833), Baf A1 (J61835), polybrene (H9268), irinotecan (I1406), all-trans-retinoic acid (R2625), and N-acetyl-l-cysteine (A9165) were all purchased from Sigma-Aldrich (USA). Anti-Top1mt–specific antibody (47 (link), 52 (link)) (Top1mt, polyclonal mouse) was a gift from Y. Pommier [Center for Cancer Research, National Cancer Institute, National Institutes of Health (NIH), USA]. Rabbit polyclonal antibodies [TDP1 (Ab4166), PGC1α (Ab54481), NRF1 (Ab34682), and mtTFAM (Ab47517)] and mouse monoclonal antibodies [COX IV (Ab14744), DRP1 (Ab56788), Mfn1 (Ab57602), and double-stranded DNA (dsDNA) (Ab27156)] were purchased from Abcam (Cambridge, USA). Mouse monoclonal antibodies [FLAG (M2) (F3165)] and rabbit polyclonal antibodies [FLAG (F7425) and LC3B (L7543)] were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse monoclonal γH2AX (05-636) antibody was purchased from Millipore, USA. Rabbit monoclonal GFP (G10362) and anti OXPHOS complex kit (457999) antibodies were purchased from Invitrogen. Mouse monoclonal actin (SC-8432) and secondary antibodies [horseradish peroxidase–conjugated anti-rabbit immunoglobulin G (IgG) or anti-mouse IgG] were purchased from Santa Cruz Biotechnology (Santa Cruz, USA). MitoTracker red (M22426), MitoTracker green (M7514), TMRM (T668), CM-H₂DCFDA (C6827), and LysoTracker red DND-99 (L7528) were obtained from Molecular Probes.

Ghosh A., Bhattacharjee S., Chowdhuri S.P., Mallick A., Rehman I., Basu S, & Das B.B. (2019). SCAN1-TDP1 trapping on mitochondrial DNA promotes mitochondrial dysfunction and mitophagy. Science Advances, 5(11), eaax9778.

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Lysosomal Enzyme Inhibitor Protocol

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E-64-d (4321-v), pepstatin A (4397), and leupeptin (4041) were purchased from the Peptide Institute (Osaka, Japan). LysoTracker Red DND-99 (L-7528) and Alexa 647–conjugated EGF (E-35351) were purchased from Molecular Probes. Bafilomycin A₁ was purchased from Wako Chemicals (Osaka, Japan). Puromycin (P8333) was purchased from Sigma-Aldrich.

Jiang P., Nishimura T., Sakamaki Y., Itakura E., Hatta T., Natsume T, & Mizushima N. (2014). The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17. Molecular Biology of the Cell, 25(8), 1327-1337.

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Fibroblast Analysis of Gaucher's Disease

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Fibroblasts of WT and type 2 GD patients were obtained from Cornell Cel Repositories (Camden, NJ, USA). Antibodies targeted for GFP (sc-9996), His-tag (sc-57598), and GAPDH (sc-25778) were all purchased from Santa Cruz Biotechnology (Dallas, TX USA). PCDGF antibody (40—3400) was procured from Invitrogen (Waltham, MA, USA). ERp57 antibody (G117) was procured from Cell Signaling Technology (Danvers, MA, USA). All fluorescence-labeled secondary antibodies used in these experiments were purchased from Jackson Immuno Research Laboratories (West Grove, PA, USA). The substrate 4-Methylumbelliferyl β-D-glucopyranoside (4-MUG, M3633) was acquired from Sigma-Aldrich (Natick, MA, USA). The dye, LysoTracker Red DND99 (L7528), and both resins, Pierce High-Capacity Endotoxin Removal Resin (2,162,373.3) and HisPur^™ Ni–NTA Resin (88,221), were all purchased from Thermo Fisher Scientific (Bridgewater, NJ, USA). The fluorescent stain, DAPI (H-1200), was acquired from VECTOR Laborato-ries (Burlingame, CA, USA). As delineated previously, the purified recombinant His-tag-PGRN protein was collected from HEK293T stable cell lines [30 (link)]. Fetal bovine serum (FBS, 16,000-044) as well as the Dulbecco’s modified Eagle’s medium (DMEM; 11,965-118) were both acquired from Gibco-BRL (Waltham, MA, USA).

Loftin C.D., Trivedi D.B., Tiano H.F., Clark J.A., Lee C.A., Epstein J.A., Morham S.G., Breyer M.D., Nguyen M., Hawkins B.M., Goulet J.L., Smithies O., Koller B.H, & Langenbach R. (2001). Failure of ductus arteriosus closure and remodeling in neonatal mice deficient in cyclooxygenase-1 and cyclooxygenase-2. Proceedings of the National Academy of Sciences of the United States of America, 98(3), 1059-1064.

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Lysosomal Enzyme Activity Assay

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All chemicals and reagents were purchased either from Sigma-Aldrich (Merck) or Thermo Fisher Scientific (Invitrogen). 4-PBA, 4-methylumbelliferyl β-D-glucopyranoside (MUD), ceapinA-7, FK506, GSK2606414, kira6, puromycin, resazurin, thapsigargin and tunicamycin were purchased from Sigma-Aldrich (Merck).
Conduritol B epoxide (CBE) was procured from Tocris Bioscience.
LysoTracker Red DND-99 (L7528) and DQ-Red BSA (D12051) were obtained from Thermo Fisher Scientific (Invitrogen). The following commercial polyclonal and monoclonal antisera were used (m, mouse; h, human and r, rat proteins): anti-LIMPII (ab16522) was from Abcam; anti-Calnexin (610523) and anti-GM130 (610822) were from BD Biosciences; anti-CHOP (2895), anti-hLAMP-1 (9091), anti-Rab5 (3547) and anti-TFEB (4240) were from Cell Signalling Technology; anti-hCD63 (H5C6) and anti-hLAMP-1 (H4A3) were from Developmental Studies Hybridoma Bank; anti-ATF6α (sc-22 799) and anti-XBP-1 (sc-7160) were from Santa Cruz Biotechnology; anti-β-actin (A5441), anti-FLAG M2 (F1804), anti-hGlucocerebrosidase (GBA/GC; G4171) and anti-TFE3 (HPA023881) from Sigma-Aldrich (Merck). All secondary antibodies were either from Invitrogen or Jackson ImmunoResearch.

Patel S., Bhatt A.M., Bhansali P, & Setty S.R.G. (2023). Pseudophosphatase STYXL1 depletion enhances glucocerebrosidase trafficking to lysosomes via ER stress. Traffic (Copenhagen, Denmark), 24(7).

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Antibody and Fluorescence Reagent Protocol

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The following antibodies were used in this study: GCase monoclonal antibody 8E4 (from J Barranger, University of Pittsburgh; 1:1000), GCase polyclonal antibody (G4171, Sigma-Aldrich, St Louis, MO, USA; 1:1000), β-actin monoclonal antibody (Sigma-Aldrich; 1:10 000), p62 monoclonal antibody (c2384-0B, BD Transduction Laboratories, Swampscott, MA, USA; 1:1000), ubiquitin polyclonal antibodies (Dako, Glostrup, Denmark and Chemicon, Temecula, CA, USA; 1:1000), α-synuclein monoclonal antibody (610787, BD Biosciences; 1:1500), α-synuclein monoclonal antibody Ab274 (1:1500),^{17 (link)} α-synuclein monoclonal antibody Ab62 (1:1000),^{17 (link)} Horse radish peroxidase (HRP)-conjugated goat anti-mouse immunoglobulin G (IgG; H+L) (172–1011, Bio-Rad Laboratories, Hercules, CA, USA; 1:3000) and HRP-conjugated goat anti-rabbit IgG (H+L) (Bio-Rad Laboratories; 1:3000).
Fluorescein-conjugated dextran (10 000 molecular weight; D-1821), TO-PRO-3 iodide (T3605) and LysoTracker Red DND-99 (L-7528) were purchased from Invitrogen (Carlsbad, CA, USA).

Bae E.J., Yang N.Y., Lee C., Lee H.J., Kim S., Sardi S.P, & Lee S.J. (2015). Loss of glucocerebrosidase 1 activity causes lysosomal dysfunction and α-synuclein aggregation. Experimental & Molecular Medicine, 47(3), e153-.

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Autophagy and Apoptosis Assays

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α-Hed(A0742) and Tax(A0177) were purchased from Chengdu Must Bio-Technology (Chengdu, China); Dulbecco’s modified Eagle’s medium (DMEM), streptomycin, penicillin, HBSS, and fetal bovine serum (FBS) were from Gibco Life Technologies (Grand Island, NY, USA); Acridine Orange (AO, A6014), and Baf (B1793) were from Sigma Biotechnology (St. Louis, MO, USA); LysoTracker Red DND-99 (L7528) was from Invitrogen (Carlsbad, CA, USA); anti-LC3B antibody (12994S), anti-p62 antibody (5114), anti-PARP antibody (9542), and cleaved-caspase 3 antibody (9662) were from Cell Signaling Technology (Boston, MA, USA); CatD antibody (SC-13985) was from Santa Cruz Biotechnology (Dallas, TX, USA). Peroxidase-labeled antibody to mouse IgG (AS003) and peroxidase-labeled antibody to rabbit IgG (AS014) were from ABclonal (Wuhan, China).
The GFP-LC3B and mCherry-GFP-LC3B plasmids were gifts from professor William KK Wu (The Chinese University of Hong Kong).

Zhan Y., Wang K., Li Q., Zou Y., Chen B., Gong Q., HO H.I., Yin T., Zhang F., Lu Y., Wu W., Zhang Y., Tan Y., Du B., Liu X, & Xiao J. (2018). The Novel Autophagy Inhibitor Alpha-Hederin Promoted Paclitaxel Cytotoxicity by Increasing Reactive Oxygen Species Accumulation in Non-Small Cell Lung Cancer Cells. International Journal of Molecular Sciences, 19(10), 3221.

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Oxidative Stress and Autophagy Modulation

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P₄ (P0130) and 17β-estradiol (E₂, E8875) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sesame oil (241002500) was purchased from Acros Organics (Leicestershire, UK). Three-methyladenine (3-MA) (M9281) and wortmannin (W1628) were purchased from Sigma-Aldrich. LysoTracker Red DND-99 (L7528), MitoTracker Red CMXRos (M7512), and non-fluorescent compound 5-(and-6)-chloromethyl-2', 7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H₂DCFDA) (C6827) were purchased from Invitrogen (Carlsbad, CA, USA). H₂O₂ was purchased from Sigma-Aldrich.

Shin H., Choi S, & Lim H.J. (2014). Relationship between reactive oxygen species and autophagy in dormant mouse blastocysts during delayed implantation. Clinical and Experimental Reproductive Medicine, 41(3), 125-131.

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Reagents and Antibodies for Cell Culture

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Regents and antibodies. Cycloheximide (5087390001), MG-132 (474791), wortmannin (W3144), 3methyladenine (M9281), brefeldin A (B5936), CDDP (PHR1624) was purchased from Sigma-Aldrich. LysoTracker Red DND-99 (L7528) and Lipofectamine 3000 (L3000015) were from Invitrogen. Anti-p53 (sc-126, Mouse mAb), anti-ATG5 (sc-133158) and anti-SQSTM1 (sc-28359) antibodies were purchased from Santa Cruz Biotechnology. Anti-ubiquitin (58395S) and anti-K63-polyubiquitin (12930S) antibodies were purchased from Cell Signaling Technology. Anti-LC3 antibody (NB100-2220) was purchased from Novus Biologicals. Anti-GAPDH antibodies (AB9132) was purchased from Chemicon. Goat anti-rabbit (Alexa Fluor 488, A-31566) and goat anti-mouse (Alexa Fluor 647, A-21242) secondary antibodies were from Invitrogen.
Cells. Cell culture reagents were purchased from Gibco (Carlsbad, CA). ES-2 and SK-BR-3 were grown in McCoy's 5A Medium. NCI-H1299 and HUVEC were grown in RPMI 1640 medium. MDA-MB-231, MIA PaCa-2 and HEK 293 were grown in DMEM medium. All mediums were supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco) except for MIA PaCa-2, and MIA PaCa-2 were supplemented with 10% fetal bovine serum (Gibco), 2.5% horse serum and 1% penicillin-streptomycin (Gibco). All cell lines were from ATCC and were cultured at 37°C under 5 % CO 2 .

Huang X., Cao Z., Qian J., Ding T., Wu Y., Zhang H., Zhong S., Wang X., Ren X., Zhang W., Xu Y., Yao G., Wang X., Yang X., Wen L, & Zhang Y. (2024). Nanoreceptors promote mutant p53 protein degradation by mimicking selective autophagy receptors. Nature nanotechnology, 19(4).

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Antibodies and Fluorescent Markers for Protein Analysis

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The following antibodies were used in this study: ATP13A2 polyclonal antibody (Abcam, ab135674, Cambridge, MA; 1:1,000), β-actin monoclonal antibody AC-15 (Sigma-Aldrich, A5441, St. Louis, MO; 1:10,000), p62 monoclonal antibody (BD Transduction Laboratories, c2384-0B, Swampscott, MA; 1:1,000), ubiquitin polyclonal antibodies (Dako, #z0458, Glostrup, Denmark, and Chemicon, Temecula, CA; 1:1,000), α-synuclein monoclonal antibody (BD Biosciences, #610787, San Diego, CA; 1:1,500), α-synuclein monoclonal antibody Ab274 (1:1,500), α-synuclein monoclonal antibody Ab62 (1:1,000), HRP-conjugated goat anti-mouse IgG (H+L) (Bio-Rad Laboratories, 172-1011, Hercules, CA; 1:3,000), and HRP-conjugated goat anti-rabbit IgG (H+L) (Bio-Rad Laboratories; 1:3,000).
Fluorescein-conjugated dextran (10,000 molecular weight; D-1821), TO-PRO-3 iodide (T3605), and LysoTracker Red DND-99 (L-7528) were purchased from Invitrogen (Carlsbad, CA).

Bae E.J., Lee C., Lee H.J., Kim S, & Lee S.J. (2014). ATP13A2/PARK9 Deficiency Neither Cause Lysosomal Impairment Nor Alter α-Synuclein Metabolism in SH-SY5Y Cells. Experimental Neurobiology, 23(4), 365-371.

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