Anti lc3b polyclonal antibody

Manufactured by Merck Group

Sourced in United Kingdom, China, Spain

The Anti-LC3B polyclonal antibody is a laboratory research tool used to detect the LC3B protein, which is associated with autophagy processes in cells. It is a purified antibody raised against a synthetic peptide derived from the human LC3B protein sequence. The antibody can be used in various immunodetection techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and localization of LC3B in biological samples.

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Lab products found in correlation

Anti erk antibody, by Cell Signaling Technology (1 mentions) Anti gapdh polyclonal antibody, by Merck Group (1 mentions) Affinipure goat anti mouse igg h l, by ZSGB-BIO (1 mentions) Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (1 mentions) Anti p62, by Abcam (1 mentions) Rapamycin, by Cell Signaling Technology (1 mentions) Lipofectaminetm rnaimax, by Thermo Fisher Scientific (1 mentions) Stealth sirna, by Thermo Fisher Scientific (1 mentions) Anti 4e bp1 antibody, by Cell Signaling Technology (1 mentions) Poweruptm sybr green master mix assay, by Thermo Fisher Scientific (1 mentions) Anti phosphorylated p70 s6 kinase, by Cell Signaling Technology (1 mentions) Anti phosphorylated ampkα thr172 antibody, by Cell Signaling Technology (1 mentions) Monoclonal anti gapdh antibody, by Merck Group (1 mentions) Pi3k inhibitor ly294002, by Cell Signaling Technology (1 mentions) Deoxycholate, by Merck Group (1 mentions) Ethd 1, by Thermo Fisher Scientific (1 mentions) 3 methyladenine, by Selleck Chemicals (1 mentions) Tris glycine sds buffer 10, by Bio-Rad (1 mentions) Tris glycine buffer 10, by Bio-Rad (1 mentions) Sterile filtered dimethyl sulfoxide, by Merck Group (1 mentions)

4 protocols using anti lc3b polyclonal antibody

Western Blot Analysis of Cardiac Proteins

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Left ventricle or cardiomyocyte whole lysates or nuclear and cytoplasmic fractions of cardiomyocytes extracted with a NE-PER Nuclear and Cytoplasmic Extraction Kit (Cat#: 78833, ThermoFisher Scientific, United States) were subject to Western blot analysis as we previously described.(Li et al., 2009b (link), Li et al., 2014 ; Qin et al., 2016 (link)) Antibodies used included anti-APG5L/ATG5 monoclonal antibody (Cat#: 3447-1, Abcam, Cambridge, United Kingdom), anti-LC3B polyclonal antibody (Cat#: L7543, Sigma-Aldrich, St. Louis, MO, United States), anti-ERK antibody (Cat#: 9101S, Cell Signaling Technology, United States), anti-angiotensinogen (Cat#: sc-7419, Santa Cruz Biotechnology Inc., Dallas, TX, United States), anti-p62 (Cat#: ab91526, Abcam, Cambridge, United Kingdom), anti-NQO1 monoclonal antibody (Cat#: sc-376023, Santa Cruz Biotechnology, Inc., Dallas, Texas, United States), anti-Nrf2 polyclonal antibody (Cat#: sc-722, Santa Cruz Biotechnology, Inc., Dallas, TX, United States), anti-GAPDH polyclonal antibody (Cat#: G9545, Sigma-Aldrich, St. Louis, MO, United States), peroxidase-conjugated AffiniPure goat anti-Mouse IgG (H+L) (Cat#: ZB2305, ZSGB-BIO, Beijing, China), and peroxidase-conjugated AffiniPure rabbit anti-goat IgG (H+L) (Cat#: ZB2306, ZSGB-BIO, Beijing, China).

Wu W., Qin Q., Ding Y., Zang H., Li D.S., Nagarkatti M., Nagarkatti P., Wang W., Wang X, & Cui T. (2021). Autophagy Controls Nrf2-Mediated Dichotomy in Pressure Overloaded Hearts. Frontiers in Physiology, 12, 673145.

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Immunofluorescent detection of LC3B

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Cells were fixed with 4% paraformaldehyde for 30 min, washed three times with PBS, and then incubated with 0.1% Triton X-100 for permeabilization. Cells were stained with anti-LC3B polyclonal antibody (Sigma, L7543) overnight at 4°C and then incubated with Alexa Fluor 488–conjugated goat anti-rabbit IgG (Abcam, ab150077) at room temperature for 1 h. Nuclei were stained with DAPI. Images were captured using a fluorescent microscope (OLYMPUS, BX53).

Huang Y.H., Sun Y., Huang F.Y., Li Y.N., Wang C.C., Mei W.L., Dai H.F., Tan G.H, & Huang C. (2017). Toxicarioside O induces protective autophagy in a sirtuin-1-dependent manner in colorectal cancer cells. Oncotarget, 8(32), 52783-52791.

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Signaling Pathway Analysis of LPA-Induced Autophagy

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Lysophosphatidic acid (LPA) (oleoyl C: 18:1) was obtained from Avanti Polar Lipids (Alabaster, AL, United States). 3-(4-[4-([1-(2-chlorophenyl)ethoxy]carbonylamino)-3-methyl-5-isoxazolyl]benzylsulfanyl)propanoic acid (Kil6425) was purchased from Sigma (St. Louis, MO, United States). PI3K inhibitor LY294002, rapamycin, anti-phosphorylated mTOR (ser2448) antibody, anti-total mTOR antibody, anti-p70s6 kinase (Thr389) antibody, anti-phosphorylated p70s6 kinase, anti-4E-BP1 antibody, anti-phosphorylated 4E-BP1(Thr37/46) antibody, and anti-phosphorylated AMPKα (Thr172) antibody were obtained from Cell Signaling Technology (Beverly, MA, United States). Anti-p62/SQSTM1 (sequestosome1) polyclonal antibody and anti-beclin1 polyclonal antibody were from Proteintech Technology (Proteintech Group, Wuhan, China), and anti-LC3B polyclonal antibody and anti-GAPDH monoclonal antibody were obtained from Sigma (St. Louis, MO, United States). Lipofectamine^TM RNAiMAX, stealth siRNA, and siRNA negative control were purchased from life Technologies (Invitrogen, Carlsbad, CA, United States). The PowerUp^TM SYBR Green Master Mix assay (Applied Biosystems, Life Technologies, Foster City, CA, United States).

Yang J., Xu J., Han X., Wang H., Zhang Y., Dong J., Deng Y, & Wang J. (2018). Lysophosphatidic Acid Is Associated With Cardiac Dysfunction and Hypertrophy by Suppressing Autophagy via the LPA3/AKT/mTOR Pathway. Frontiers in Physiology, 9, 1315.

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Autophagy Modulation Experimental Protocol

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Chemical salts (NaCl, KCl, MgSO4, KPO4, sodium pyruvate, sodium lactate, CaCl₂, NaHCO₃, Na₃VO₄), HEPES, EGTA, EDTA, deoxycholate, Triton X-100, glucose, bovine serum albumin (BSA), formaldehyde, phosphate buffered saline (PBS), chloroquine diphosphate salt (Ref# C6628), sterile-filtered dimethyl sulfoxide (DMSO; Ref# D2650), E-64d (Aloxistatin) (Ref# E8640), and anti-LC3B polyclonal antibody (Ref# L7543) were purchased from Sigma-Aldrich (Madrid, Spain). 3-Methyladenine (Ref# S2767) was purchased from Selleck Chemicals (Planegg, Germany). Anti-Beclin-1 pAb (Ref# PD017Y), Anti-Atg16 pAb (Ref# PM040Y), and a positive control for anti-LC3 (Ref# PM036-PNY) were purchased from MBL International (Woburn, MA, USA). EthD-1 and the anti-rabbit IgG antibody labeled with Alexa 546 were obtained from Molecular Probes (Thermo Fisher Scientific, Madrid, Spain). Anti-rabbit IgG horseradish peroxidase (HRP)-conjugated secondary antibody and enhanced chemiluminescence detection reagents were from Pierce Protein Biology (Thermo Fisher Scientific, Madrid, Spain). A complete, EDTA-free, protease inhibitor cocktail was from Roche Diagnostics (Barcelona, Spain). Bradford reagent, Tris/Glycine/SDS buffer (10×), and Tris/Glycine buffer (10×) were from Bio-Rad (Madrid, Spain). STF-62247 (Ref# 189497) was from Merck Millipore (Darmstadt, Germany).

Aparicio I.M., Rojo-Domínguez P., Castillejo-Rufo A., Peña F.J, & Tapia J.A. (2023). The Autophagy Marker LC3 Is Processed during the Sperm Capacitation and the Acrosome Reaction and Translocates to the Acrosome Where It Colocalizes with the Acrosomal Membranes in Horse Spermatozoa. International Journal of Molecular Sciences, 24(2), 937.

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