LC3A Antibody

Western Blot: LC3A Antibody [NB100-2331] - Analysis of heart tissue lysates from mice which were subjected or not to 48 hours of starvation. The signal was developed using ECL method and this LC3 antibody was found to detect both forms of LC3, i.e. LC3-I and LC3-II. As expected, the levels of LC3-II form were higher in the heart tissue lysates from starved mice.

Western Blot: LC3A Antibody [NB100-2331] - Detection of HRP conjugated autophagic LC3 in mouse ES cell lysate. The atg5-/- lane (ES cells, cultured to form embryonic bodies, that are deficient in conversion of LC3-1 to LC3-11) demonstrates the specificity of NB 100-2331, as there is no detection of LC3-11. Photo courtesy of Dr. Beth Levine, UT Southwestern Medical Center.

Western Blot: LC3A Antibody [NB100-2331] - Analysis in human cell lysates.

Immunocytochemistry/Immunofluorescence: LC3A Antibody [NB100-2331] - Analysis in HeLa cells using anti-LC3 antibody (red). Nuclei were counterstained with DAPI (blue).

Western Blot: LC3A Antibody [NB100-2331] - LC3A expression in Mouse macrophage cell line RAW 264.7. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: LC3A Antibody [NB100-2331] - Left panel shows untreated HeLa cells. Right panel shows HeLa cells that were treated with 50 uM CQ overnight. Cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti-LC3A antibody at 5 ug/mL overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: LC3A Antibody [NB100-2331] - Staining in mouse meniscus and cartilage. Image from verified customer review.

Western Blot: LC3A Antibody [NB100-2331] - Analysis in human brain lyaste.

Immunocytochemistry/Immunofluorescence: LC3A Antibody [NB100-2331] - Analysis in PFA fixed NIH/3T3 cells using anti-LC3A antibody. Image from verified customer review.

Immunohistochemistry: LC3A Antibody [NB100-2331] - Staining of human brain, cerebral cortex, cell processes in gray matter.

Immunohistochemistry-Paraffin: LC3A Antibody [NB100-2331] - Analysis in mouse renal tissue. Image from verifed customer review.

Simple Western: LC3A Antibody [NB100-2331] - Image shows a specific band for LC3 in 0.5 mg/mL of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt, Ca, Fi, Pl, Ze, Bv
• Applications: WB, Simple Western, ELISA, Flow, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, SB
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

LC3A Antibody Summary

• Immunogen: A synthetic peptide made to an internal portion of the human LC3 protein sequence (between residues 25-121). [Uniprot: Q9H492].
• Localization: LC3-I is cytoplasmic. LC3-II binds to the autophagic membranes.
• Marker: Autophagosome Marker
• Specificity: This antibody detects both LC3A and LC3B.
• Predicted Species: Bovine (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: MAP1LC3A
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot
  • Simple Western 1:50
  • ELISA
  • Flow Cytometry
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:100 - 1:300
  • Immunohistochemistry 1:200 - 1:400
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200 - 1:400
  • Immunoprecipitation 20 ug / 500 ug of lysate
  • Southern Blot
• Application Notes: Western blot bands are seen at ~19 kDa, representing LC3-I, and ~17 kDa, representing LC3-II. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. In ICC, cytoplasmic staining was observed in HeLa cells. Use in immunocytochemistry (PMID: 21545732) and Immunohistochemistry-Paraffin (PMID: 26571030) reported in scientific literature. Use in Flow cytometry reported in scientific literature ( PMID: 24419333). Use in ELISA reported in scientific literature (PMID: 20930550). Use in Southern blot reported in scientific literature (PMID: 21262964). Use in immunoblotting reported in scientific literature (PMID: 28253371). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
• Control:

  LC3B Lysate (NBL1-12844)
  Brain Lysate (NB820-59657)
  Brain Lysate (NB820-59177)
  Neuro2a Lysate (NBP2-19017)
  Neuro2a Lysate (NBP2-49688)
  HeLa Lysate (NBP2-49689)

Reactivity Notes

Fish reactivity reported in scientific literature (PMID: 25522711). Canine reactivity reported in scientific literature (PMID: 26179070). Plant reactivity reported in scientific literature (PMID: 27861739).

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for LC3A Antibody

• Apg8
• APG8a
• Apg8p3
• ATG8E
• Autophagy-related protein LC3 A
• Autophagy-related ubiquitin-like modifier LC3 A
• LC3
• LC3A
• lc3-i, ii
• MAP1A/1B light chain 3 A
• MAP1ALC3
• MAP1ALC3MAP1A/MAP1B LC3 A
• MAP1BLC3MAP1A/MAP1B light chain 3 A
• MAP1LC3A
• microtubule-associated protein 1 light chain 3 alphaMAP1 light chain 3-like protein 1
• microtubule-associated proteins 1A/1B light chain 3
• microtubule-associated proteins 1A/1B light chain 3A
• MLP3A

Background

LC3 (microtubule-associated protein light chain 3), the most studied autophagy biomarker, was originally identified as a subunit of microtubule-associated proteins 1A and 1B (MAP1LC3) and was later found to contain similarity to yeast protein Apg8/Aut7/Cvt5. Distributed ubiquitously in eukaryotes, LC3 is expressed as 3 splice variants/isoforms (LC3A, LC3B and LC3C) which undergo post-translational processing, wherein, the unprocessed form of LC3 is proteolytically cleaved by Atg4 protease to form LC3-I with carboxyterminal exposed glycine. During autophagy, this exposed glycine of LC3-I is conjugated by Atg7 (an E1-like activity), Atg3 (an E2-like conjugating activity) and by Atg12-Atg5-Atg16L multimers (E3-like ligase activity) to phosphatidylethanolamine (PE) moiety for generating LC3-II. The lipophilic character of PE group facilitates LC3-II insertion into autophagosomes membranes, and as a result LC3-II is degraded when autophagosomes fuse with lysosomes to form autolysosomes for lysus of intra-autophagosomal components by lysosomal hydrolases. Conversion of LC3I to LC3II when correlated with autophagosome numbers is considered as the best marker of autophagy because LC3-II is the only well-characterized protein which specifically localize to autophagic structures throughout autophagy (from phagophore to lysosomal degradation). LC3 is a great tool in research as autophagy is implicated in numerous physiological/pathological processes including responses to exercise/aging, cancer, metabolic and neurodegenerative disorders, and cardiovascular/pulmonary diseases.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for LC3A Antibody (NB100-2331)(220)

Publications using NB100-2331

• Maier L, Kallenberger S, Jechow K et al. Unraveling mitotic protein networks by 3D multiplexed epitope drug screening bioRxiv [PMID: 30104419] (ICC/IF, Human)
• Karanasios E, Ktistakis NT. Studying Autophagy: List of Useful Antibodies Produced via a Community Effort. Autophagy at the Cell, Tissue and Organismal Level. 2016 Apr 24
• Hollins SL, Brock L, Barreto R et al. A Rodent Model of Anxiety: The Effect of Perinatal Immune Challenges on Gastrointestinal Inflammation and Integrity. Neuroimmunomodulation. Nov 9 2018 [PMID: 30415249] (IHC, Rat)
• Hollins SL, Brock L, Barreto R et al. A Rodent Model of Anxiety: The Effect of Perinatal Immune Challenges on Gastrointestinal Inflammation and Integrity. Neuroimmunomodulation. Nov 9 2018 [PMID: 30415249] (IHC, Rat)
• Hanemaaijer E, Panahi M, Swaddiwudhipong N et al. Autophagy-lysosomal defect in human CADASIL vascular smooth muscle cells. European Journal of Cell Biology. Oct 1 2018 [PMID: 30392756] (ICC/IF, Human)
• Ozturk DG, kocak M, Akcay A et al. MITF-MIR211 axis is a novel autophagy amplifier system during cellular stress. Autophagy. Oct 5 2018 [PMID: 30290719] (WB, Human)
• Schaffner I, Minakaki G, Khan MA et al. FoxO Function Is Essential for Maintenance of Autophagic Flux and Neuronal Morphogenesis in Adult Neurogenesis. Neuron Sep 19 2018 [PMID: 30197237] (WB, Mouse)
• Battista RA, Resnati M, Facchi C et al. Autophagy mediates epithelial cancer chemoresistance by reducing p62/SQSTM1 accumulation PLoS ONE Aug 1 2018 [PMID: 30067838] (WB, Human)
• Kustermann M, Manta L, Paone C et al. Loss of the novel Vcp (valosin containing protein) interactor Washc4 interferes with autophagy-mediated proteostasis in striated muscle and leads to myopathy in vivo. Autophagy Aug 16 2018 [PMID: 30010465] (WB, Zebrafish)
• Lu JM, Dong WM, He HC et al. Autophagy induced by overexpression of DCTPP1 promotes tumor progression and predicts poor clinical outcome in prostate cancer Int. J. Biol. Macromol. Jun 3 2018 [PMID: 29874556] (Human)
• Saera-Vila A, Kish PE, Kahana A. Autophagy in Zebrafish Extraocular Muscle Regeneration Methods Mol. Biol. May 24 2018 [PMID: 29797006] (Zebrafish)
• Chmielewska M, Dedukh D, Haczkiewicz K et al. The programmed DNA elimination and formation of micronuclei in germ line cells of the natural hybridogenetic water frog Pelophylax esculentus Sci Rep May 18 2018 [PMID: 29777142] (ICC/IF)
• Humphrey SA. DNA damage and oxidative stress induced-p53 activity in astrocytes causes growth arrest. 2012 University of Western Ontario.
• Lim H, Lim YM, Kim KH et al. A novel autophagy enhancer as a therapeutic agent against metabolic syndrome and diabetes. Nat Commun Apr 12 2018 [PMID: 29650965] (WB, Human)
• Anunobi R, Boone BA, Cheh N et al. Extracellular DNA promotes colorectal tumor cell survival after cytotoxic chemotherapy. J. Surg. Res. Jun 1 2018 [PMID: 29605400] (WB, ICC/IF)
• Fusco C, Mandriani B, Di Rienzo M et al. TRIM50 regulates Beclin 1 proautophagic activity. Biochim Biophys Acta Mol Cell Res 2018 Jun [PMID: 29604308]
• Jeong AL, Ka HI, Han S et al. Oncoprotein CIP2A promotes the disassembly of primary cilia and inhibits glycolytic metabolism. EMBO Rep. Feb 28 2018 [PMID: 29491003] (WB)
• Piacentini M, Baiocchini A, Del Nonno F, Melino G. Non-alcoholic fatty liver disease severity is modulated by transglutaminase type 2. Cell Death Dis. 2018 Feb 15 [PMID: 29449533] (WB, Mouse)
• Inomata Y, Nagasaka S, Miyate K, Goto Y. Bcl-2-associated athanogene 3 (BAG3) is an enhancer of small heat shock protein turnover via activation of autophagy in the heart. Biochem. Biophys. Res. Commun. 2018 Feb 19 [PMID: 29409895] (WB, Mouse)
• Festa BP, Chen Z, Berquez M et al. Impaired autophagy bridges lysosomal storage disease and epithelial dysfunction in the kidney Nat Commun 2018 Jan 11 [PMID: 29323117] (WB, Zebrafish)
• Watanabe S, Komine O, Endo F et al. Intracerebroventricular administration of Cystatin C ameliorates disease in SOD1-linked amyotrophic lateral sclerosis mice. J. Neurochem. 2017 Dec 28 [PMID: 29282717] (WB, Mouse)
• Palucci I, Matic I, Falasca L et al. Transglutaminase type 2 plays a key role in the pathogenesis of Mycobacterium tuberculosis infection. J. Intern. Med. 2017 Dec 04 [PMID: 29205566] (Mouse)
• Minakaki G, Menges S, Kittel A et al. Autophagy inhibition promotes SNCA/alpha-synuclein release and transfer via extracellular vesicles with a hybrid autophagosome-exosome-like phenotype. Autophagy 2017 Dec 04 [PMID: 29198173] (WB, Mouse)
• Jiang MY, Han ZD, Li W et al. Mitochondrion-associated protein peroxiredoxin 3 promotes benign prostatic hyperplasia through autophagy suppression and pyroptosis activation Oncotarget. 2017 Oct 06 [PMID: 29113303] (WB, Human)
• Desseille C, Deforges S, Biondi O et al. Specific Physical Exercise Improves Energetic Metabolism in the Skeletal Muscle of Amyotrophic-Lateral-Sclerosis Mice. Frontiers in Molecular Neuroscience. 2017 Oct 20 [PMID: 29104532] (WB, Mouse)
• Theofilas P, Ehrenberg AJ, Nguy A et al. Probing the correlation of neuronal loss, neurofibrillary tangles, and cell death markers across the Alzheimer's disease Braak stages: a quantitative study in humans. Neurobiol. Aging. 2017 Sep 20 [PMID: 29031088] (ICC/IF, IHC, Human)
• Kaverina N, Borovjagin AV, Kadagidze Z et al. Astrocytes promote progression of breast cancer metastases to the brain via a KISS1-mediated autophagy. Autophagy. 2017 Oct 05 [PMID: 28981380] (ICC/IF)
• Solanas G, Peixoto FO, Perdiguero E et al. Aged Stem Cells Reprogram Their Daily Rhythmic Functions to Adapt to Stress Cell 2017 Aug 10 [PMID: 28802040] (Mouse)
• Benitez BA, Sands MS. Primary fibroblasts from CSPalpha mutation carriers recapitulate hallmarks of the adult onset neuronal ceroid lipofuscinosis Sci Rep 2017 Jul 24 [PMID: 28740222] (WB, Human)
• Nazim UM, Moon JH, Lee YJ et al. PPARy activation by troglitazone enhances human lung cancer cells to TRAIL-induced apoptosis via autophagy flux. Oncotarget Apr 18 2017 [PMID: 28460464] (WB, Human)
• Bae SY, Byun S, Bae SH et al. TPT1 (tumor protein, translationally-controlled 1) negatively regulates autophagy through the BECN1 interactome and an MTORC1-mediated pathway. Autophagy. Feb 15 2017 [PMID: 28409693] (WB, Mouse)
• Yue F, Li W, Zou J et al. Spermidine prolongs lifespan and prevents liver fibrosis and hepatocellular carcinoma by activating MAP1S-mediated autophagy. Cancer Res. Apr 6 2017 [PMID: 28386016] (WB, Human)
• Lopez A, Lee SE, Wojta K et al. A152T tau allele causes neurodegeneration that can be ameliorated in a zebrafish model by autophagy induction. Brain. Feb 9 2017 [PMID: 28334843] (WB, Fish)
• Mattoscio D, Casadio C, Miccolo C et al. Autophagy regulates UBC9 levels during viral-mediated tumorigenesis. PLoS Pathog. Mar 1 2017 [PMID: 28253371] (IB, Human)
• Garcia-Prat L, Munoz-Canoves P, Martinez-Vicente M. Monitoring Autophagy in Muscle Stem Cells. Methods Mol. Biol. Mar 1 2017 [PMID: 28247355] (Mouse)
• Schulze U, Vollenbroker B, Kuhnl A et al. Cellular vacuolization caused by overexpression of the PIKfyve-binding deficient Vac14(L156R) is rescued by starvation and inhibition of vacuolar-ATPase. Biochim. Biophys. Acta. 2017 Feb 16 [PMID: 28216340] (ICC/IF)
• Suzuki J, Nakajima W, Suzuki H et al. Chaperone-mediated autophagy promotes lung cancer cell survival through selective stabilization of the pro-survival protein, MCL1. Biochem. Biophys. Res. Commun. Dec 7 2016 [PMID: 27939888] (WB, Human)
• Lu Y, Liu XF, Liu TR et al. Celecoxib exerts antitumor effects in HL-60 acute leukemia cells and inhibits autophagy by affecting lysosome function. Biomed. Pharmacother. Nov 21 2016 [PMID: 27884749] (Human)
• Lee S-, Kang J-, Kim S- et al. Afzelin ameliorates Dgalactosamine and lipopolysaccharideinduced fulminant hepatic failure by modulating mitochondrial quality control and dynamics. British Journal of Pharmacology. Nov 15 2016 [PMID: 27861739] (WB, Mouse, Plant)
• Shingu T, Ho AL, Yuan L et al. Qki deficiency maintains stemness of glioma stem cells in suboptimal environment by downregulating endolysosomal degradation. Nat. Genet. Nov 14 2016 [PMID: 27841882] (WB, Mouse)
• Jiang C, Jin S, Jiang Z, Wang J. Inhibitory effects of silibinin on proliferation and lung metastasis of human high metastasis cell line of salivary gland adenoid cystic carcinoma via autophagy induction. Onco Targets Ther. Nov 8 2016 [PMID: 27822066] (Mouse)
• Zhang D, Pan J, Xiang X et al. Protein Kinase Cdelta Suppresses Autophagy to Induce Kidney Cell Apoptosis in Cisplatin Nephrotoxicity. J. Am. Soc. Nephrol. Oct 31 2016 [PMID: 27799485]
• Song X, Xie Y, Kang R et al. FANCD2 protects against bone marrow injury from ferroptosis. Biochem. Biophys. Res. Commun. Nov 18 2016 [PMID: 27773819]
• Liu P, Jin H, Guo Z et al. Silver nanoparticles outperform gold nanoparticles in radiosensitizing U251 cells in vitro and in an intracranial mouse model of glioma. Int J Nanomedicine. Oct 19 2016 [PMID: 27757033] (ICC/IF, Human)
• Benvenuto M, Mattera R, Masuelli L et al. Gossypol induces apoptosis and autophagy in head and neck carcinoma cell lines and inhibits the growth of transplanted salivary gland cancer cells in BALB/c mice. Int J Food Sci Nutr Sep 27 2016 [PMID: 27670669]
• Morell C, Bort A, Vara-Ciruelos D et al. Up-Regulated Expression of LAMP2 and Autophagy Activity during Neuroendocrine Differentiation of Prostate Cancer LNCaP Cells PLoS ONE Sep 15 2016 [PMID: 27627761] (WB, Human)
• Yamada E, Okada S, Bastie CC et al. Fyn phosphorylates AMPK to inhibit AMPK activity and AMP-dependent activation of autophagy. Oncotarget Sep 8 2016 [PMID: 27626315]
• Mo Zt, Li Wn, Zhai Yr, Gong Qh. Icariin Attenuates OGD/R-Induced Autophagy via Bcl-2-Dependent Cross Talk between Apoptosis and Autophagy in PC12 Cells. Evid Based Complement Alternat Med. Jul 25 2016 [PMID: 27610184] (WB)
• Caccamo A, Ferreira E, Branca C, Oddo S p62 improves AD-like pathology by increasing autophagy. Mol. Psychiatry 2016 Aug 30 [PMID: 27573878]
• Saini NK, Baena A, Ng TW et al. Suppression of autophagy and antigen presentation by Mycobacterium tuberculosis PE_PGRS47. Nat Microbiol. Aug 26 2016 [PMID: 27562263] (WB, Mouse)
• Tan KP, Ho MY, Cho HC et al. Fucosylation of LAMP-1 and LAMP-2 by FUT1 correlates with lysosomal positioning and autophagic flux of breast cancer cells. Cell Death Dis. Aug 26 2016 [PMID: 27560716] (ICC/IF, Human)
• Sun X, Sun L, Zhao Y et al. MAVS maintains mitochondrial homeostasis via autophagy. Cell Discov. Aug 23 2016 [PMID: 27551434] (IP, WB, Human)
• Sabha N, Volpatti JR, Gonorazky H et al. PIK3C2B inhibition improves function and prolongs survival in myotubular myopathy animal models. J. Clin. Invest. Sep 1 2016 [PMID: 27548528] (WB, Mouse)
• Hong JM, Kim SJ, Lee SM. Role of necroptosis in autophagy signaling during hepatic ischemia and reperfusion. Toxicol. Appl. Pharmacol. Aug 10 2016 [PMID: 27521978] (WB)
• Jiuju Du, Wencheng Zhu, Li Yang et al. Reduction of polyethylenimine-coated iron oxide nanoparticles induced autophagy and cytotoxicity by lactosylation. Oxford Academics 2016 Jun 12 [PMID: 27482464] (WB, Mouse)
• Granato M, Zompetta C, Vescarelli E et al. HCV derived from sera of HCV-infected patients induces pro-fibrotic effects in human primary fibroblasts by activating GLI2. Sci Rep. Aug 1 2016 [PMID: 27476557] (WB, Human)
• Wu HM, Wang J, Zhang B et al. CpG-ODN promotes phagocytosis and autophagy through JNK/P38 signal pathway in Staphylococcus aureus-stimulated macrophage. Life Sci. Jul 27 2016 [PMID: 27476088] (WB, Mouse)
• Saera-Vila A, Kish PE, Louie KW et al. Autophagy Regulates Cytoplasmic Remodeling During Cell Reprogramming in a Zebrafish Model of Muscle Regeneration. Autophagy. Jul 28 2016 [PMID: 27467399] (WB, Zebrafish)
• Suzuki K, Yamaguchi A, Yamanaka S et al. Accumulated a-synuclein affects the progression of GM2 gangliosidoses Exp. Neurol. 2016 Jul 21 [PMID: 27453479]
• Lee HY, Kim J, Quan W et al. Autophagy deficiency in myeloid cells increases susceptibility to obesity-induced diabetes and experimental colitis. Autophagy Jun 23 2016 [PMID: 27337687] (WB)
• Erbil S, Oral O, Mitou G et al. RACK1 is an Interaction Partner of ATG5 and a Novel Regulator of Autophagy. J. Biol. Chem. Jun 20 2016 [PMID: 27325703] (WB)
• Hossini AM, Quast AS, Plotz M et al. PI3k/AkT Signaling Pathway Is Essential for Survival of Induced Pluripotent Stem Cells. PLoS ONE. May 4 2016 [PMID: 27138223] (WB, Human)
• Piras A, Collin L, Gruninger F et al. Autophagic and lysosomal defects in human tauopathies: analysis of post-mortem brain from patients with familial Alzheimer disease, corticobasal degeneration and progressive supranuclear palsy. Acta Neuropathol Commun 2016 Mar 02 [PMID: 26936765] (WB)
• Pema M, Drusian L, Chiaravalli M et al. mTORC1-mediated inhibition of polycystin-1 expression drives renal cyst formation in tuberous sclerosis complex. Nat Commun 2016 Mar 02 [PMID: 26931735] (WB)
• Jacob F, Yonis AY, Cuello F et al. Analysis of Tyrosine Kinase Inhibitor-Mediated Decline in Contractile Force in Rat Engineered Heart Tissue. PLoS One 2016 Feb 03 [PMID: 26840448] (WB)
• Buhler A, Kustermann M, Bummer T et al. Atrogin-1 Deficiency Leads to Myopathy and Heart Failure in Zebrafish. Int J Mol Sci 2016 Jan 30 [PMID: 26840306] (WB)
• Li W, Zou J, Yue F et al. Defects in MAP1S-mediated autophagy cause reduction in mouse lifespans especially when fibronectin is overexpressed. Aging Cell. 2016 Jan 10 [PMID: 26750654] (WB, Mouse)
• Garcia-Prat L, Martinez-Vicente M, Perdiguero E et al. Autophagy maintains stemness by preventing senescence. Nature. 2016 Jan 07 [PMID: 26738589] (WB, ICC/IF, Mouse)
• Altuntas S, Rossin F, Marsella C et al. The transglutaminase type 2 and pyruvate kinase isoenzyme M2 interplay in autophagy regulation. Oncotarget 2015 Dec 29 [PMID: 26702927] (WB)
• Duarte-Silva S, Silva-Fernandes A, Neves-Carvalho A et al. Combined therapy with m-TOR-dependent and -independent autophagy inducers causes neurotoxicity in a mouse model of Machado-Joseph disease. Neuroscience. 2015 Nov 19 [PMID: 26601773] (WB, Mouse)
• Song K, Hu W, Yue F et al. Transforming Growth Factor TGFbeta Increases Levels of Microtubule-Associated Protein MAP1S and Autophagy Flux in Pancreatic Ductal Adenocarcinomas. PLoS ONE. 2015 Nov 17 [PMID: 26571030] (IHC-P, Human)
• Muhammad E, Levitas A, Singh SR et al. PLEKHM2 mutation leads to abnormal localization of lysosomes, impaired autophagy flux and associates with recessive dilated cardiomyopathy and left ventricular noncompaction. Hum Mol Genet 2015 Dec 20 [PMID: 26464484] (Human)
• Xiong R, Zhou W, Siegel D et al. A Novel Hsp90 Inhibitor Activates Compensatory Heat Shock Protein Responses and Autophagy and Alleviates Mutant A53T alpha-Synuclein Toxicity. Mol Pharmacol 2015 Dec [PMID: 26405178] (Human)
• Grossi V, Lucarelli G, Forte G et al. Loss of STK11 expression is an early event in prostate carcinogenesis and predicts therapeutic response to targeted therapy against MAPK/p38. Autophagy 2015 Sep 21 [PMID: 26391455] (WB, Human)
• Daniels CMLP, Paffenroth E, Austin EV, Glebov K. Lithium Decreases Glial Fibrillary Acidic Protein in a Mouse Model of Alexander Disease. PloS one 2015 Sep 17 [PMID: 26378915] (WB, Mouse)
• Chen Q, Yue F, Li W et al. Potassium Bisperoxo (1,10-phenanthroline) Oxovanadate (bpV(phen)) Induces Apoptosis and Pyroptosis and Disrupts the P62-HDAC6 Interaction to Suppress the Acetylated Microtubule-dependent Degradation of Autophagosomes. J. Biol. Chem. 2015 Sep 11 [PMID: 26363065] (WB, Human, Mouse)
• Zhen Y, Li W. Impairment of autophagosome-lysosome fusion in the buff mutant mice with the VPS33A(D251E) mutation. Autophagy. 2015 Aug 11 [PMID: 26259518] (WB, Mouse)
• Pomilio C, Pavia P, Gorojod RM et al. Glial alterations from early to late stages in a model of alzheimer's disease: Evidence of autophagy involvement in Abeta internalization. Hippocampus. 2015 Aug 01 [PMID: 26235241] (IHC, Mouse)
• Irimia JM, Tagliabracci VS, Meyer CM et al. Muscle glycogen remodeling and glycogen phosphate metabolism following exhaustive exercise of wild type and laforin knockout mice. J. Biol. Chem. 2015 Jul 27 [PMID: 26216881] (WB, Mouse)
• Kabak YB, Sozmen M, Yarim M et al. Immunohistochemical detection of autophagy-related microtubule-associated protein 1 light chain 3 (LC3) in the cerebellums of dogs naturally infected with canine distemper virus Biotech Histochem. 2015 Jul 16 [PMID: 26179070] (IHC-P, Canine)
• Gorojod RM, Alaimo A, Porte Alcon S et al. The autophagic- lysosomal pathway determines the fate of glial cells under manganese- induced oxidative stress conditions Free Radic. Biol. Med. 2015 Jul 07 [PMID: 26163003] (IHC-Fr, Rat)
• Chia-Wei Lin, Ming-Shiou Jan, Jung-Hua Steven Kuo. Exploring MicroRNA Expression Profiles Related to the mTOR Signaling Pathway in Mouse Embryonic Fibroblast Cells Treated with Polyethylenimine. Mol Pharm. 2015 Aug 3 [PMID: 26158199]
• Nabissi M, Morelli MB, Amantini C et al. Cannabidiol stimulates Aml-1a-dependent glial differentiation and inhibits glioma stem-like cells proliferation by inducing autophagy in a TRPV2-dependent manner. Int. J. Cancer. 2015 Apr 22 [PMID: 25903924] (WB, Human)
• Wang T, Martin S, Papadopulos A et al. Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a J. Neurosci. 2015 Apr 15 [PMID: 25878289] (WB, ICC/IF, Rat)
• Ulasov IV, Shah N, Kaverina NV et al. Tamoxifen improves cytopathic effect of oncolytic adenovirus in primary glioblastoma cells mediated through autophagy Oncotarget 2015 Feb 28 [PMID: 25738357] (WB, Human)
• Wei Y, An Z, Zou Z et al. The stress-responsive kinases MAPKAPK2/MAPKAPK3 activate starvation-induced autophagy through Beclin 1 phosphorylation Elife. 2015 Feb 19 [PMID: 25693418] (WB, Human)
• Zou J, Li W, Misra A et al. Viral Restriction Factor Tetherin Prevents Leucine-rich Pentatricopeptide Repeat Containing Protein (LRPPRC) from Association with Beclin 1 and B-cell CLL/lymphoma 2 (Bcl-2) and Enhances Autophagy and Mitophagy J. Biol. Chem. 2015 Jan 28 [PMID: 25631043] (WB, Human)
• Nuschke A, Rodrigues M, Stolz DB et al. Human mesenchymal stem cells/multipotent stromal cells consume accumulated autophagosomes early in differentiation. Stem Cell Res Ther. 2014 Dec 17 [PMID: 25523618] (WB, Human)
• Wilson WN, Baumgarner BL, Watanabe WO et al. Effects of resveratrol on growth and skeletal muscle physiology of juvenile southern flounder. Comp. Biochem. Physiol., Part A Mol. Integr. Physiol. 2014 Dec 16 [PMID: 25522711] (WB, Fish)
• Varga M, Fodor E, Vellai T. Autophagy in zebrafish. Methods. 2014 Dec 09 [PMID: 25498006] (WB, Zebrafish)
• Cicchini M, Chakrabarti R, Kongara S et al. Autophagy regulator BECN1 suppresses mammary tumorigenesis driven by WNT1 activation and following parity. Autophagy 2014 Oct 30 [PMID: 25483966] (IHC)
• Gan M, Moussaud S, Jiang P, Mclean Pj. Extracellular ATP induces intracellular alpha-synuclein accumulation via P2X1 receptor-mediated lysosomal dysfunction Neurobiology of Aging et al. 2014 Nov 05 [PMID: 25480524] (WB, Human)
  
  Details:
  LC3/MAP1LC3A antibody used for WB on lysates of S1S2 cells (derived from founder H4/Tetoff cells) and differentiated Lund human mesencephalic/Luhmes cells treated or not with 3 mM ATP or 200 nM bafilomycin A1 for 48 hours (Fig. 6B and Fig. 6E).
• Carlisi D, Lauricella M, D'Anneo A et al. The synergistic effect of SAHA and Parthenolide in MDA-MB231 breast cancer cells J. Cell. Physiol. 2014 Nov 05 [PMID: 25370819] (WB, Human)
  
  Details:
  LC3 antibody used for WB on MDA-MB231 breast cancer cells treated or not with suberoylanilide hydroxamic acid (SAHA, an histone deacetylase inhibitor), sesquiterpene lactone Parthenolide (Pn) and bafilomycin A1 (BafA1, a powerful inhibitor of the autophagic flux). WB data is shown in Figure 3b.
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• Li J, Rohailla S, Gelber N et al. MicroRNA-144 is a circulating effector of remote ischemic preconditioning. Basic Res. Cardiol. 2014 Sep 01 [PMID: 25060662] (WB, Mouse)
  
  Details:
  LC3 antibody used for WB in hearts from mice injected with PBS, miR-Co or miR-144 (Figure 2b).
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• Zou J, Yue F, Jiang X et al. Mitochondrion-associated protein LRPPRC suppresses the initiation of basal levels of autophagy via enhancing Bcl-2 stability. Biochem J 2013 Jul 3 [PMID: 23822101] (WB, Human)
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• Chen JJ, Long ZJ, Xu DF et al. Inhibition of autophagy augments the anticancer activity of a -mangostin in chronic myeloid leukemia cells. Taylor Francis. 2014 Mar [PMID: 23734655] (WB, Human)
• Xiong R, Siegel D, Ross D. The activation sequence of cellular protein handling systems after proteasomal inhibition in dopaminergic cells. Chem Biol Interact 2013 May 14 [PMID: 23684743] (WB, Rat)
• Palmio J, Evila A, Chapon f et al. Hereditary myopathy with early respiratory failure: occurrence in various populations. J Neurol Neurosurg Psychiatry 2013 Apr 19 [PMID: 23606733]
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• Chen MC, Chen CH, Liu YN et al. TW01001, a novel piperazinedione compound, induces mitotic arrest and autophagy in non-small cell lung cancer A549 cells. Cancer Lett 2013 Apr 6 [PMID: 23567646] (WB, Human)
• Martin S, Harper CB, May LM et al. Inhibition of PIKfyve by YM-201636 Dysregulates Autophagy and Leads to Apoptosis-Independent Neuronal Cell Death PLoS One 2013 [PMID: 23544129] (WB, Mouse)
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• Drerup CM, Nechiporuk AV. JNK-Interacting Protein 3 Mediates the Retrograde Transport of Activated c-Jun N-Terminal Kinase and Lysosomes. PLoS Genet 2013 Feb [PMID: 23468645] (WB, IHC, ICC/IF, Zebrafish)
• Chiou SK, Hoa N, Ge L, Jadus MR. Nutrient Availability Alters the Effect of Autophagy on Sulindac Sulfide-Induced Colon Cancer Cell Apoptosis. Gastroenterol Res Pract. 2012 [PMID: 23431290] (WB, Human)
• Xie X, White EP, Mehnert JM. Coordinate Autophagy and mTOR Pathway Inhibition Enhances Cell Death in Melanoma. PLoS One 2013 [PMID: 23383069] (WB, Human)
• Teplova I, Lozy F, Price S et al. ATG proteins mediate efferocytosis and suppress inflammation in mammary involution. Autophagy 2013 Apr 1 [PMID: 23380905] (IHC, Mouse)
• Shoji-Kawata S, Sumpter R, Leveno M et al. Identification of a candidate therapeutic autophagy-inducing peptide. Nature 2013 Feb 14 [PMID: 23364696] (WB, Human)
• Choi BY, Jang BG, Kim JH et al. Copper/zinc chelation by clioquinol reduces spinal cord white matter damage and behavioral deficits in a murine MOG-induced multiple sclerosis model. Neurobiol Dis 2013 Jan 26 [PMID: 23360710] (IHC-Fr, ICC/IF, WB, Mouse)
• Jiang TF, Zhang YJ, Zhou HY et al. Curcumin Ameliorates the Neurodegenerative Pathology in A53T alpha-synuclein Cell Model of Parkinson's Disease Through the Downregulation of mTOR/p70S6K Signaling and the Recovery of Macroautophagy. J Neuroimmune Pharmacol 2013 Jan 17 [PMID: 23325107] (WB, ICC/IF, Human)
• Balaburski GM, Leu JI, Beeharry N et al. A Modified HSP70 Inhibitor Shows Broad Activity as an Anticancer Agent. Mol Cancer Res 2013 Mar [PMID: 23303345] (WB, Human)
• Momma K, Noguchi S, Malicdan MC et al. Rimmed vacuoles in becker muscular dystrophy have similar features with inclusion myopathies PLoS One 2012 [PMID: 23251671] (IHC, ICC/IF, Human)
• Jena P, Mohanty S, Mohanty T et al. Azurophil granule proteins constitute the major mycobactericidal proteins in human neutrophils and enhance the killing of mycobacteria in macrophages PLoS One 2012 [PMID: 23251364] (ICC/IF, Human)
• Martinet W, Schrijvers DM, Timmermans JP et al. Immunohistochemical analysis of macroautophagy: recommendations and limitations. Autophagy 2013 Mar [PMID: 23242143] (Mouse)
• Sulzmaier FJ, Li Z, Nakashige ML et al. AD65, GAD67, and GABAT Immunostaining in Human Brain and Apparent GAD65 Loss in Alzheimer's Disease PLoS One 2012 [PMID: 23144724] (WB, Human)
• Nakagawa S, Nishihara K, Inui KI, Masuda S. Involvement of autophagy in the pharmacological effects of the mTOR inhibitor everolimus in acute kidney injury Eur J Pharmacol 2012 Sep 26 [PMID: 23022334] (WB, IHC-Fr, ICC/IF, Rat)
• Lin CW, Jan MS, Kuo JH et al. Protective role of autophagy in branched polyethylenimine (25K)- and poly(L-lysine) (30-70K)-induced cell death. Eur J Pharm Sci. 2012 Dec 18 [PMID: 23000135]
• Cyrino LT, Araujo AP, Joazeiro PP et al. In vivo and in vitro Leishmania amazonensis infection induces autophagy in macrophages Tissue Cell 2012 Aug 30 [PMID: 22939777] (WB, ICC/IF, Mouse)
• He C, Sumpter R Jr, Levine B. Exercise induces autophagy in peripheral tissues and in the brain Autophagy. 2012 Oct [PMID: 22892563] (WB, Mouse)
• Joselin AP, Hewitt SJ, Callaghan SM et al. ROS-dependent regulation of Parkin and DJ-1 localization during oxidative stress in neurons Hum Mol Genet 2012 Aug 23 [PMID: 22872702] (ICC/IF, Mouse)
• Ramos FJ, Chen SC, Garelick MG et al. Rapamycin Reverses Elevated mTORC1 Signaling in Lamin A/C-Deficient Mice, Rescues Cardiac and Skeletal Muscle Function, and Extends Surviva Sci Transl Med. 2012 Jul 25 [PMID: 22837538]
• Garcia-Garcia A, Rodriguez-Rocha H, Tseng MT et al. E2F-1 lacking the transcriptional activity domain induces autophagy Cancer Biol Ther 2012 Sep 1 [PMID: 22825328]
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• Grkovic S, O'Reilly VC, Han S et al. IGFBP-3 binds GRP78, stimulates autophagy and promotes the survival of breast cancer cells exposed to adverse microenvironments Oncogene 2012 Jul 2 [PMID: 22751133] (WB, Human)
• Eijiro Yamada, Claire C Bastie, Hiroshi Koga, Yichen Wang, Ana Maria Cuervo, Jeffrey E Pessin. Mouse Skeletal Muscle Fiber-Type-Specific Macroautophagy and Muscle Wasting Are Regulated by a Fyn/STAT3/Vps34 Signaling Pathway. Cell Reports, 10.1016/j.celrep.03.014. 2012 [PMID: 22745922] (WB, Mouse)
• Claerhout S, Dutta B, Bossuyt W et al. Abortive Autophagy Induces Endoplasmic Reticulum Stress and Cell Death in Cancer Cells PLoS One 2012 [PMID: 22745748] (WB, Human)
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• Liang X, De Vera ME, Buchser WJ et al. Inhibiting Systemic Autophagy during Interleukin 2 Immunotherapy Promotes Long-term Tumor Regression Cancer Res 2012 Jun 1 [PMID: 22472122] (IHC-Fr, ICC/IF, WB, Mouse)
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• Martinet W, Verheye S, De Meyer I, Timmermans JP, Schrijvers DM, Van Brussel I, Bult H, De Meyer GR. Everolimus triggers cytokine release by macrophages: rationale for stents eluting everolimus and a glucocorticoid. Arterioscler Thromb Vasc Biol;32(5):1228-35. 2012 May. [PMID: 22345170] (WB, Mouse)
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• Mostowy S, Sancho-Shimizu V, Hamon M et al. p62 and NDP52 target Intracytosolic Shigella and Listeria to different autophagy pathways. J Biol Chem. 2011 Jun 6. [PMID: 21646350]
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• Fazio EN, Dimattia GE, Chadi SA et al. Stanniocalcin 2 alters PERK signaling and reduces cellular injury during cerulein induced pancreatitis in mice. BMC Cell Biol;12:17. 2011 May 5. [PMID: 21545732]
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• Rossin F, D'Eletto M, Macdonald D, Farrace MG, Piacentini M. TG2 transamidating activity acts as a reostat controlling the interplay between apoptosis and autophagy. Amino Acids;42(5):1793-802. 2012 May. [PMID: 21479826] (WB, Mouse)
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• Uddin MN, Nishio N, Ito S, Suzuki H, Isobe K. Autophagic activity in thymus and liver during aging. Age (Dordr);34(1):75-85. 2012 Feb. [PMID: 21387084] (IHC, Mouse)
• Tang Y-C, Williams BR, Siegel JJ, Amon A. Identification of Aneuploidy-Selective Antiproliferation Compounds. Cell;144(4):499-512. 2011 Feb 18. [PMID: 21315436]
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• Hsu HS, Huang PI, Chang YL et al. Cucurbitacin I inhibits tumorigenic ability and enhances radiochemosensitivity in nonsmall cell lung cancer-derived CD133-positive cells. Cancer. Jul. 2011 [PMID: 21225866] (WB, Human)
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• Yoon S, Woo SU, Kang JH et al. STAT3 transcriptional factor activated by reactive oxygen species induces IL6 in starvation-induced autophagy of cancer cells. Autophagy 2010 Nov [PMID: 20930550] (ELISA, Human)
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Reviews for LC3A Antibody (NB100-2331) (18)

Average Rating: 4.3

(Based on 18 reviews)

We have 18 reviews tested in 4 species: Human, Mouse, Mouse and Human, Other.

reviewed by: Anonymous WB Mouse and Human 11/25/2017

Summary

• Application: Western Blot
• Sample Tested: mouse hepatocytes and HepG2 cells
• Species: Mouse and Human
• Lot: AC-1

reviewed by: Anonymous WB Mouse 08/07/2017

Summary

• Application: Western Blot
• Sample Tested: Mouse macrophage cell line RAW 264.7
• Species: Mouse
• Lot: H6-NB100-2331

reviewed by: jennifer guadagno WB Mouse 10/06/2015

Summary

• Application: Western Blot
• Sample Tested: Mouse Neuro2A, Mouse Cortical Neurons whole cell lysate
• Species: Mouse
• Lot: R-4

Blocking

• Blocking Details: 10% non-fat dry milk in TBS-T (0.05% Tween) for 1hr at room temperature

Primary Anitbody

• Dilution Ratio: 1:1000

Secondary Antibody

• Secondary Description: Goat-anti rabbit HRP
• Secondary Concentration: 1:5000 in 3% milk in TBS-T (0.05%)

Details

• Detection Notes: Clarity ECL, Biorad Chemidoc imager, exposed for 1 min, Neg control= untreated cells, Pos Control= rapamycin

Comments

• Comments: This antibody works very well. Very strong clear signal.

reviewed by: Anonymous WB Human 01/12/2015

Summary

• Application: Western Blot
• Sample Tested: human glioblastoma
• Species: Human

Blocking

• Blocking Details: Li-Cor blockin bufffer

Primary Anitbody

• Dilution Ratio: 2 ug/ml, overnight, room temperature, TBST buffer

Secondary Antibody

• Secondary Description: anti-IgG secondary antibodies, Conjugation: IRDye® 800CW
• Secondary Manufacturer Cat#: Li-Cor
• Secondary Concentration: per vendor recomendations

Details

• Detection Notes: Odyssey Li-Cor

Comments

• Comments: Please site our paper:Tamoxifen improves cytopathic effect of oncolytic adenovirus in primary glioblastoma cells mediated through autophagy." just accepted for Oncotarget.

reviewed by: Anonymous WB Mouse 12/12/2014

Summary

• Application: Western Blot
• Sample Tested: See PMID 22892563
• Species: Mouse

Blocking

• Blocking Details: See PMID 22892563

Details

• Detection Notes: See PMID 22892563

Comments

• Comments: Published in PMID: 22892563

reviewed by: Anonymous IF Mouse 11/25/2014

Summary

• Application: Immunofluorescence
• Sample Tested: NIH3T3, HeLa, HEK293T, N2a neuroblastoma, Q7 striatal, Sy5Y neuroblastoma
• Species: Mouse

Blocking

• Blocking Details: 1% BSA + 1%milk

Primary Anitbody

• Dilution Ratio: 1:100 for IF

Secondary Antibody

• Secondary Description: Cy5 conjuagated anti-rabbit
• Secondary Concentration: 1:500

Details

• Detection Notes: Images taken by Zeiss Axiovert microscope
• Fixation Details: PFA Fixed, 0.1% Tween20 permeabilized, PBS, 1hr primary incubation at RT
• Wash Description: PBS 3x washes, 2 mins each

reviewed by: Merissa Olmer IHC-P 10/04/2014

Summary

• Application: Immunohistochemistry-Paraffin
• Lot: S-1
• Comments: These tissue samples were fixed with zinc buffered formalin.

reviewed by: Ilya Ulasov WB Human 06/30/2014

Summary

• Application: Western Blot
• Sample Tested: human tumor cells
• Species: Human
• Lot: q-1

Blocking

• Blocking Details: #917-40000, LI-COR

Primary Anitbody

• Dilution Ratio: in agreement with vendor recommendation

Secondary Antibody

• Secondary Description: goat-anti-rabbit
• Secondary Manufacturer Cat#: Li-COR

Details

• Detection Notes: Odyssey Imaging System (Model #1866, Li-COR Biosciences

reviewed by: Anonymous WB Human 06/20/2014

Summary

• Application: Western Blot
• Species: Human
• Pub Med ID: 24735980
• File: View PDF

reviewed by: Kumsal Tekirdag WB Human 04/02/2014

Summary

• Application: Western Blot
• Species: Human
• Lot: S-2
• Special Applications: Gives slighter LC3-I band compared to LC3II. Stronger binding to LC3II. High level of background at first usage in BSA solution.
• Pub Med ID: 24358205
• File: View PDF

reviewed by: Nirmala Parajuli IHC Mouse 08/13/2012

Summary

• Application: Immunohistochemistry
• Sample Tested: Renal tissue (paraffin block)
• Species: Mouse
• Lot: N
• Comments: Antibody is good for immunohistochemistry. But, since this is polyclonal, each lot has to be tested for concentration of primary antibody to get desired results.

Blocking

• Blocking Details: Protein block, Dako, 20 min RT

Primary Anitbody

• Dilution Ratio: 1:10,000, 4dC, ON, 0.1%BSA plus0.05% skim milk

Secondary Antibody

• Secondary Description: Goat anti-rabbit-HRP
• Secondary Manufacturer Cat#: DAKO, #K4011
• Secondary Concentration: DAKO kit

Details

• Detection Notes: DAB, 5 min
• Fixation Details: 10% NBF, 10mM Sodium Citrate buffer pH6.0
• Wash Description: TBS-T, 5 min X 4

Comments

• Comments: Antibody is good for immunohistochemistry. But, since this is polyclonal, each lot has to be tested for concentration of primary antibody to get desired results.

reviewed by: Anonymous WB Human 12/28/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human
• Comments: Good antibody!

Blocking

• Blocking Details: Blocking Buffer: 10% non-fat milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight

Secondary Antibody

• Secondary Description: Secondary Ab: rabbit

Details

• Detection Notes: Detection Method: HRP

Comments

• Comments: Good antibody!

reviewed by: Anonymous WB Human 11/21/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: 5% Milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab Dilution Ratio: 1:5000

Details

• Detection Notes: Detection Method: ECL-HRP

reviewed by: Anonymous WB Human 11/21/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: 2% Skim Milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab: Goat anti-rabbit HRP

Details

• Detection Notes: Detection Method: ECL

reviewed by: Anonymous WB Mouse 10/24/2011

Summary

• Application: Western Blot
• Sample Tested: Mouse
• Species: Mouse
• Comments: The antibody worked well!

Blocking

• Blocking Details: Blocking Buffer: 5% Milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: 16 hours

Secondary Antibody

• Secondary Description: Secondary Ab: anti-mouse HRP antibody

Details

• Detection Notes: Detection Method: ECL

Comments

• Comments: The antibody worked well!

reviewed by: Seung-Hyun Ro WB Mouse 07/08/2010

Summary

• Application: Western Blot
• Sample Tested: 3T3-L1 adipocyte, Sample Amount: 30ug
• Species: Mouse
• Lot: H2
• Comments: I used in for 3T3-L1 mouse adipocyte and it is working great.

Blocking

• Blocking Details: Blocking Buffer: 5% Milk in PBST, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1:1000, Incubation Dilution Buffer: PBST, Incubation Time: 2 hours, Incubation Temp: room temperature

Secondary Antibody

• Secondary Description: Secondary Ab: Rabbit, Secondary Ab Dilution Ratio: 1:3000
• Secondary Manufacturer Cat#: Santa Cruz sc-2077

Details

• Detection Notes: Detection Method: ECL - perkin elmer, Exposure Time: 1 minute, Positive Control: Actin, Specific Bands: 15, 18 kDa

Comments

• Comments: I used in for 3T3-L1 mouse adipocyte and it is working great.

reviewed by: Anonymous WB Other 11/05/2009

Summary

• Application: Western Blot
• Sample Tested: lysed cell lines, Sample Amount: 20 ug
• Species: Other
• Lot: J

Blocking

• Blocking Details: Blocking Buffer: TBS 0.1% Tween20 5% NFDM, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Dilution Concentration: 0.002, Incubation Dilution Buffer: TBS 0.1% Tween20 5% NFDM, Incubation Time: 1 hr, Incubation Temp: RT

Secondary Antibody

• Secondary Description: Secondary Ab: Goat anti-Rabbit HRP, Secondary Ab Dilution Ratio: 1/4000

Details

• Detection Notes: Detection Method: Super Signal West Dura, Exposure Time: 1 minute, Specific Bands: 19 and 17kDa doublet

reviewed by: Anonymous WB Human 02/18/2009

Summary

• Application: Western Blot
• Sample Tested: Human cancer cells, Sample Amount: 80ug
• Species: Human
• Lot: SS

Blocking

• Blocking Details: Blocking Buffer: 5% Milk in PBST, Blocking Time: 30 minutes, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Details

• Detection Notes: Detection Method: Licor, Exposure Time: 2 minutes, Specific Bands: 17 and 19 kDa

FAQs for LC3A Antibody (NB100-2331). (Showing 1 - 3 of 3 FAQs).

1. May we ask if it is possible to perform IF to stain LC3-I and LC3-II separately with two different fluorescent colors?
   • Yes, it is possible to perform IF stain for LC3-I and LC3-II separately with two different fluorescent colors! You will have to use two different primary antibodies and in order to avoid any potential background/cross reactivity issues, I would suggest that you employ conjugated primary antibodies for the testing. 1. Our LC3I antibody (NBP1-78964) has been designed to specifically detect the cytosolic form of the LC3 protein which is actually LC3 I (Note: LC3-II binds to the autophagic membranes). 2. There is not even a single antibody to our knowledge that would exclusively detect the LC3 II form, and you would have to detect LC3II/ autophagic membranes form with an antibody which detects LC3 I/LC3II together. Therefore you may opt second antibody from one of the followings: LC3 Antibody (NB100-2220), LC3 Antibody (NB100-2331), LC3 Antibody (NBP1-19167). All of these mentioned catalog #s come with different options for their conjugated forms and you may select appropriate conjugated forms for performing the IF staining using our explained criteria.
2. I am interested in detecting the LC3 protein in the sea anemone Aiptasia. I have used your LC3 antibody (NB100-2331) and got a band in my Western blot at around 15 kD, which seems reasonable. However I would like to know how likely it is that your antibody binds to the LC3 protein of Aiptasia? Unfortunately I could not find the protein sequence against which the LC3 antibody was raised to and am hoping you could help me. The protein sequence for the Aiptasia LC3 is: MGDNNVLSYKPFKQRKSFVSRRDEVAGIRAKFPSKVPVIVERYHKERDLPLLDKTKFLVPQELTMSQFVTIIRNRMSLSS TQAFYLIVNNKSLASMSMTMAELYREEKDEDGFLYMVYASQEMFGCNS
   • In comparing the sequences of the human LC3 and Aiptasia proteins, and it seems as though there is very low homology. There is only 62% sequence similarity, and we generally don't recommend antibodies for novel species unless they have at least 85%. There is a chance that it will work, but we cannot guarantee it.
3. Do you have any data or reason to believe the your LC3 antibody (NB100-2331) has a higher affinity to LC3-II than to LC3-I?
   • No, we do not have any data or reason to believe that NB100-2331 has a higher affinity to LC3-II than to LC3-I. This antibody was raised using a synthetic peptide made to an internal portion of the human LC3 protein sequence (between residues 25-121) as an immunogen and is expected to detect both LC3-I as well as LC3-II, and their signal will depend upon their respective amounts being present in your samples at the time of detection.

Bioinformatics

• Gene Symbol: MAP1LC3A