Lc3 l7543

Manufactured by Merck Group

Sourced in United States

The LC3 (L7543) is a laboratory equipment product from Merck Group. It is a liquid chromatography system designed for analytical and preparative separations. The core function of the LC3 is to perform high-performance liquid chromatography (HPLC) analysis and purification. The product specifications and detailed technical details are available upon request.

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Lab products found in correlation

Tempol, by Merck Group (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Tetramethylrhodamine methyl ester, by Thermo Fisher Scientific (1 mentions) Triclosan, by Merck Group (1 mentions) Anti p62, by Cell Signaling Technology (1 mentions) Anti p ampk, by Cell Signaling Technology (1 mentions) Anti p stat3, by Cell Signaling Technology (1 mentions) Mitotracker, by Thermo Fisher Scientific (1 mentions) Fluo 3 am, by Thermo Fisher Scientific (1 mentions) Bcl 2 ab196495, by Abcam (1 mentions) Calam, by Thermo Fisher Scientific (1 mentions) Anti stat3, by Cell Signaling Technology (1 mentions) Anti ampk, by Cell Signaling Technology (1 mentions) Mitosox, by Thermo Fisher Scientific (1 mentions) Rapamycin, by Cayman Chemical (1 mentions) Sc 25792, by Santa Cruz Biotechnology (1 mentions) Hydroxychloroquine h0915, by Merck Group (1 mentions) Visionworks software, by Analytik Jena (1 mentions) Biospectrum 810 imaging system, by Analytik Jena (1 mentions) Cleaved caspase 3 9661, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

L7543 (102 citations) Anti-LC3 (L7543; (4 citations) Polyclonal antibodies against LC3 (L7543) (2 citations)

6 protocols using lc3 l7543

Triclosan-Induced Oxidative Stress and STAT3 Inhibition

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Triclosan was obtained from Shanghai Baidi Biody-Bio Co., Ltd. Hoechst, Cal-AM, Eth-1, Fluo-3/AM, mito-Tracker, mito-SOX, tetramethylrhodamine methyl ester (TMRM) and DAPI were purchased from Thermo Fisher Scientific, Inc. Dihydroethidium dye was purchased from the Beyotime Institute of Biotechnology. Tempol, 3-MA and acetylcysteine (NAC) were purchased from Sigma Aldrich; Merck KGaA. Tempol (0.5 and 1 mM) is a radical scavenger that was used to test the effect of ROS levels on cytotoxicity induced by Triclosan (20 µM) in the lactate dehydrogenase (LDH) release assay. S3I-201 was purchased from EMD Millipore. S3I-201 (10 and 20 µM) is a STAT3 inhibitor that was used to detect the effect of STAT3 activity change on cytotoxicity induced by Triclosan (20 µM) in the LDH release assay. Anti-p-STAT3 (Y705, #9131, 1:1,000), anti-STAT3 (#4904, 1:1,000), anti-p-AMPK (Thr172, #2535, 1:1,000), anti-AMPK (#2532, 1:1,000) and anti-p62 (#88588, 1:1,000) were purchased from Cell Signaling Technology, Inc., Bcl-2 (ab196495, 1:1,000) antibody was purchased from Abcam and LC3 (L7543, 1:1,000) antibody was purchased from Sigma Aldrich; Merck KGaA.

Jin J., Chen N., Pan H., Xie W., Xu H., Lei S., Guo Z., Ding R., He Y, & Gao J. (2020). Triclosan induces ROS-dependent cell death and autophagy in A375 melanoma cells. Oncology Letters, 20(4), 73.

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Garcinia subelliptica Leaf Extract Analysis

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The ethanol extract of the leaf of Garcinia subelliptica Merr. (catalog # FBM124-035) was purchased from the Korea Research Institute of Bioscience and Biotechnology (Daejeon, Korea). Doxorubicin (D1515), E-64 (E8640), pepstatin A (P4265), and hydroxychloroquine (H0915) were purchased from Sigma-Aldrich (St. Louis, MO, USA), and rapamycin (#13346) was from Cayman (Ann Arbor, MI, USA). Antibodies against PARP (#9532), mTOR (#2972), phosphor-mTOR (S2448, #2971), ULK1 (#8054), phospho-ULK1(S757, #6888), and phospho-ULK (S317, #12753) were procured from Cell signaling (Danvers, MA, USA). Anti-p62/SQSTM1 antibody was obtained from Abcam (Cambridge, UK). Antibodies against β-actin (sc-477,778) and AMPKα (sc-25,792) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and LC3 (L7543) was from Sigma-Aldrich.

Kim K.H., Lee J.Y., Li W.Y., Lee S., Jeong H.S., Choi J.Y, & Joo M. (2021). The ethanol extract of Garcinia subelliptica Merr. induces autophagy. BMC Complementary Medicine and Therapies, 21, 280.

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Western Blot Analysis of Protein Lysates

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After treatment, spheroids were lysed in cold RIPA buffer (NONIDET-P40 1%, NaDOC 0.5%, SDS 1%, in PBS). The concentration of total protein was evaluated with a colorimetric assay (DC protein assay from Bio-Rad, Hercules, CA, USA). 25 μg of cell lysates were loaded in reducing conditions (0.2 M Tris, pH 6.8, 5% SDS, 3% glycerol, 0.01% bromophenol blue and 200 mM DTT). After separation in SDS-PAGE (7.5 to 15% acrylamide) and transfer to PVDF (Immobilon, Millipore, Billerica, MA, USA), membranes were blocked with a protein-free TBS blocking buffer (Pierce, Rockford, IL, USA) and gently agitated with antibodies diluted in 5% non-fat dry milk or 5% BSA, as appropriate, at 4°C overnight. Secondary antibodies were from Amersham (Piscataway, NJ, USA). Chemiluminescence was detected by the enhanced SuperSignal West Pico Substrate (Pierce, Rockford, IL, USA) with a Biospectrum 810 imaging system (UVP, Upland, CA, USA). LC3 (#L7543) and tubulin (#T-6074) antibodies were from Sigma-Aldrich (St. Louis, MO, USA). P-AKT^Ser473 (#4060), P-S6K^Thr389 (#9234), P-ERK (#4370), Bid (#2002), ATG5 (#12994), ATG7 (#8558) and cleaved caspase 3 (#9661) antibodies were from Cell Signaling (Danver MA, USA). Bim (#559685) antibody was from BD Pharmingen (San Jose, CA, USA). Densitometry analysis was performed with VisionWorks software (UVP, Upland, CA, USA).

Barbone D., Follo C., Echeverry N., Gerbaudo V.H., Klabatsa A., Bueno R., Felley-Bosco E, & Broaddus V.C. (2015). Autophagy Correlates with the Therapeutic Responsiveness of Malignant Pleural Mesothelioma in 3D Models. PLoS ONE, 10(8), e0134825.

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Mitochondrial Regulation of Autophagy

Cited 1 time

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Chloroquine diphosphate (CQ, Sigma, C6628). Tris [(1-benzyl-1H-1,2,3-triazol-4-yl) methyl] amine (TBTA), Tris (2-carboxyethyl) phosphine (TCEP), CuSO₄ were purchased from Sigma. Rhodamine-azide was obtained from Jinglan Co. (Guangzhou, China) and biotin-azide was purchased from Thermo Fisher Scientific. The ART-probe (ART-P) was synthesized as previous description [20] (link). MitoSOX™ Red Mitochondrial Superoxide Indicator (M36008), JC-1 Mitochondrial Membrane Potential Dye (65-0851-38), MitoTracker™ Red (M7513), Pacific Blue™ Annexin V Apoptosis Detection Kit (A35122), ThiolTracker™ Violet (glutathione, T10095) were purchased from Thermo Fisher Scientific.
The following antibodies were used: autophagy-related gene 7 (ATG7) (2631), cytochrome c oxidase Ⅳ (COX Ⅳ) (4850), dynamin-related protein 1 (DRP1) (8570), phospho-DRP1 (Ser616) (3455), heat shock protein 60 (HSP60) (12165), lysosomal-associated membrane protein 1 (LAMP1) (9091), mitofusin 1 (MFN-1) (14739), MFN-2 (11925), Dynamin-like 120 kDa protein (OPA-1) (67589), PINK1 (6946), Parkin (4211), SQSTM1/P62 (5114), Ubiquitin (3936), were purchased from Cell Signaling Technology. α-tubulin (sc-23948), translocase of outer mitochondrial membrane 20 (TOMM20) (sc-11415), translocase of the inner membrane 23 (TIM23) (sc-514463) were purchased from Santa Cruz Biotechnology. LC3 (L7543) was purchased from Sigma.

Zhang J., Sun X., Wang L., Wong Y.K., Lee Y.M., Zhou C., Wu G., Zhao T., Yang L., Lu L., Zhong J., Huang D, & Wang J. (2018). Artesunate-induced mitophagy alters cellular redox status. Redox Biology, 19, 263-273.

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Protein Modulation and Detection Assay

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MG-132, cycloheximide, chloroquine, and DMSO were purchased from Sigma-Aldrich. Only chloroquine was solubilized in Milli-Q water. The following antibodies were used: anti-SMN (1:2000, 2F1, Cell Signaling Technologies); anti-p53 (PAb1801, Ref); p21 (F-5, Santa Cruz); anti-ubiquitin (FL-76, Santa Cruz); p62 (P0067, Sigma-Aldrich); LC3 (L7543, Sigma-Aldrich); anti-tubulin (DM1α, 1:4000, Sigma-Aldrich); and anti-β-actin (AC-74, 1:4000, Sigma-Aldrich).

Rietz A., Li H., Quist K.M., Cherry J.J., Lorson C.L., Burnett B., Kern N.L., Calder A.N., Fritsche M., Lusic H., Boaler P.J., Choi S., Xing X., Glicksman M.A., Cuny G.D., Androphy E.J, & Hodgetts K.J. (2017). Discovery of a Small Molecule Probe That Post-Translationally Stabilizes the Survival Motor Neuron Protein for the Treatment of Spinal Muscular Atrophy. Journal of medicinal chemistry, 60(11), 4594-4610.

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Immunoblotting of Acetylation Markers

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For whole-cell extract, cells were lysed in buffer containing 0.5% NP-40 (Roche, 11754599001), 100 mM Tris-HCl (pH 7.4), 300 mM NaCl and complemented with protease and phosphatase inhibitors [27] , AA (10 μM), and HDAC inhibitor cocktail (Santa-Cruz, sc-362323). Proteins were resolved by SDS-gel electrophoresis, and blots probed with antibodies against acetylated-K (Ac-K) (No. 9441), DRP1 (D6C7) (No. 8570), H3 (No. 9715), acetyl-H3K14 (No. 4318, Ac-H3K14 (D4B9), No. 7627), PCAF (C14G9) (No. 3378), SIRT1 (D1D7) (No. 9475), pSIRT1 (serine 47) (No. 2314), and acetyl-α-tubulin (K40) (Ac-TubK40, No. 3971) from cell signaling, acetyl-H4Ser1K5K8K12 (G-2) (Ac-H4K5K8K12, sc-393472), HDAC6 (H-300) (sc-11420), hMOF (G-12) (sc-271691), p300 (F-4) (sc-48343), TOM20 (F-10) (sc-17764), α-tubulin (TU-02) (sc-8035) from Santa Cruz, GAPDH (Millipore, NG1740950), LC3 (L7543), and SIRT1 (S5322) from Sigma-Aldrich, LONP1 (Proteintech, 15440-1-AP), subunit IVof cytochrome c oxidase (COX IV, Abcam, ab14744), PINK1 (Novus biologicals, BC100-494), and TIP60 (Calbiochem, DR1041).

Yakhine-Diop S.M.S., Niso-Santano M., Rodríguez-Arribas M., Gómez-Sánchez R., Martínez-Chacón G., Uribe-Carretero E., Navarro-García J.A., Ruiz-Hurtado G., Aiastui A., Cooper J.M., López de Munaín A., Bravo-San Pedro J.M., González-Polo R.A, & Fuentes J.M. (2019). Impaired Mitophagy and Protein Acetylation Levels in Fibroblasts from Parkinson's Disease Patients. Molecular neurobiology, 56(4).

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