LC3A Antibody

Western Blot: LC3A Antibody [NB100-2331] - High autophagosome concentration is consumed during early immortalized human mesenchymal stem cell differentiation. Immortalized human mesenchymal stem cells were differentiated under osteogenic conditions (see Materials and methods) and assayed for changes in LC3I and LC3II during a 72-hour window. Cells were differentiated under standard conditions (top) or with addition of 5 uM rapamycin (middle) or 5 nM bafilomycin (bottom) for the first 3 hours of differentiation to modulate autophagy. Immunoblots were performed for LC3 at the indicated time points to assess autophagosome degradation via relative changes in LC3II (lower band; 17 kDa). Studies were repeated three times with similar trends seen consistently. Image collected and cropped by CiteAb from the following publication (//stemcellres.com/content/5/6/140), licensed under a CC-BY licence.

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 in mouse meniscus and cartilage. Image from verified customer review.

Western Blot: LC3A Antibody [NB100-2331] - Autophagy mobilized during differentiation of immortalized human mesenchymal stem cells (ihMSCs). Cultured ihMSCs were stimulated to undergo osteogenesis to form osteoblasts for 30 days at 21% and 4% oxygen. LC3I and LC3II levels were assessed via immunoblot every 10 days and normalized to beta-actin to derive average LC3II band densities. Image collected and cropped by CiteAb from the following publication (//stemcellres.com/content/5/6/140), licensed under a CC-BY licence.

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

Western Blot: LC3A Antibody [NB100-2331] - This LC3A antibody Image shows 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. LC3A and LC3B. As expected, the levels of LC3B 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.

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

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: LC3A Antibody [NB100-2331] - Rapamycin increases autophagy in brains of PDAPP mice. Representative epifluorescent (c200x) image of hippocampal CA1 in control- and rapamycin-fed transgenic PDAPP mice stained with an anti-LC3 antibody. An increase in LC3-immunoreactive puncta was observed in CA1 projections of transgenic PDAPP mice following rapamycin administration. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0009979) licensed under a CC-BY licence.

Immunohistochemistry: 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, IHC-WhMt
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

LC3A Antibody Summary

• Immunogen: This LC3A Antibody was prepared from 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 LC3A 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
  • Immunohistochemistry Whole-Mount
• 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 Immunohistochemistry-Whole mount reported in scientific literature (PMID: 31783118). 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 Overexpression Lysate
  Mouse Brain Whole Tissue Lysate (Adult Whole Normal)
  Human Brain Whole Tissue Lysate (Adult Whole Normal)
  Neuro2a Whole Cell Lysate
  Neuro2a Chloroquine Treated / Untreated Cell Lysate
  HeLa Chloroquine Treated / Untreated Cell Lysate

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
• MAP1 light chain 3-like protein 1
• MAP1A/1B light chain 3 A
• MAP1A/MAP1B LC3 A
• MAP1A/MAP1B light chain 3 A
• MAP1ALC3
• MAP1BLC3
• MAP1LC3A
• Microtubule-associated protein 1 light chain 3 alpha
• microtubule-associated proteins 1A/1B light chain 3
• microtubule-associated proteins 1A/1B light chain 3A
• MLP3A

Background

Human Microtubule-associated Protein 1A/1B Light Chain 3A (MAP1LC3A), also called LC3A for short, is a 121 amino acid (aa) protein with a theoretic molecular weight of ~14 kDa. LC3A belongs to the LC3 subfamily of Autophagy-related 8 (Atg8) proteins, which also includes LC3B and LC3C (1). The process of autophagy is the bulk degradation of proteins and organelles. Whether these three proteins have distinct roles in autophagy remains unclear, but they do have unique subcellular expression (2). Specifically, LC3A shows perinuclear and nuclear localization (2). The Atg8 family members share a similar structure of two amino-terminal alpha-helices and a ubiquitin-like core but are unique in aa sequence (1). LC3 utilizes a ubiquitin-like conjugation system to covalently bind to phosphatidylethanolamine (PE), also called lipidation, which is mediated by a series of steps (1, 3). Briefly, unprocessed, cytosolic LC3 (pro-LC3) is cleaved by the cysteine protease Atg4 to expose a c-terminal glycine (Gly) residue (LC3-I); the Gly is activated by E1-like enzyme Atg7, transferred to E2-like enzyme Atg3, and an E3-like complex facilitates the conjugation of LC3 with PE (LC3-II), incorporating it into the phagophore membrane during autophagosome formation (1, 3). The recruitment of LC3 to the phagophore is thought to mediate membrane elongation and closure (1, 3). Additionally, LC3 plays a role in recruiting cargo (protein aggregates, pathogens, and organelles) to autophagosomes and delivery for lysosomal degradation (1).

The process of autophagy is associated with a variety of diseases including neurodegenerative diseases, neuromuscular, tumorigenesis, and viral and bacterial infections (4). LC3 is a useful marker of autophagy in both healthy and diseased cells (4). Interestingly, LC3A has two variants (v1 and v2) which differ in N-terminal sequence due to the varying transcriptional start sites (5). One particular study found that LC3Av1, but not v2 or LC3B, was silenced in various cancer cell lines due to aberrant DNA methylation and re-expression of LC3Av1 in LC3Av1-silenced cells inhibited tumor growth, where overall findings suggest a possible tumor-suppressive role (5).

Alternative names for LC3A include Apg8, APG8a, ATG8E, Autophagy-related protein LC3 A, Autophagy-related ubiquitin-like modifier LC3 A, MAP1A/1B light chain 3 A, microtubule-associated proteins 1A/1B light chain 3, and MLP3A.

References

1. Shpilka, T., Weidberg, H., Pietrokovski, S., & Elazar, Z. (2011). Atg8: an autophagy-related ubiquitin-like protein family. Genome biology. https://doi.org/10.1186/gb-2011-12-7-226

2. Koukourakis, M. I., Kalamida, D., Giatromanolaki, A., Zois, C. E., Sivridis, E., Pouliliou, S., Mitrakas, A., Gatter, K. C., & Harris, A. L. (2015). Autophagosome Proteins LC3A, LC3B and LC3C Have Distinct Subcellular Distribution Kinetics and Expression in Cancer Cell Lines. PloS one. https://doi.org/10.1371/journal.pone.0137675

3. Weidberg, H., Shvets, E., & Elazar, Z. (2011). Biogenesis and cargo selectivity of autophagosomes. Annual review of biochemistry. https://doi.org/10.1146/annurev-biochem-052709-094552

4. Tanida, I., Ueno, T., & Kominami, E. (2004). LC3 conjugation system in mammalian autophagy. The international journal of biochemistry & cell biology. https://doi.org/10.1016/j.biocel.2004.05.009

5. Schaaf, M. B., Keulers, T. G., Vooijs, M. A., & Rouschop, K. M. (2016). LC3/GABARAP family proteins: autophagy-(un)related functions. FASEB journal : official publication of the Federation of American Societies for Experimental Biology. https://doi.org/10.1096/fj.201600698R

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)(244)

Publications using NB100-2331

• Humphrey SA. DNA damage and oxidative stress induced-p53 activity in astrocytes causes growth arrest. 2012 University of Western Ontario.
• Zhang R, Varela M, Forn-CunI G et al. Deficiency in the autophagy modulator Dram1 exacerbates pyroptotic cell death of Mycobacteria-infected macrophages Cell Death Dis Apr 24 2020 [PMID: 32332700] (WB, Zebrafish)
• Fedele A, Proud C Chloroquine and bafilomycin A mimic lysosomal storage disorders and impair mTORC1 signalling. Biosci Rep. 2020 Apr 30 [PMID: 32285908] (WB, Human)
• Li W, Dai Y, Shi B et Al. LRPPRC sustains Yap-P27-mediated cell ploidy and P62-HDAC6-mediated autophagy maturation and suppresses genome instability and hepatocellular carcinomas Oncogene Mar 16 2020 [PMID: 32203162] (Mouse)
• Szekerczes T, Gogl A, Illyes I et Al. Autophagy, Mitophagy and MicroRNA Expression in Chronic Hepatitis C and Autoimmune Hepatitis Pathol. Oncol. Res. Mar 2 2020 [PMID: 32124227] (IHC-P, Human)
• Cai K Altered expression levels of autophagy-associated proteins during exercise preconditioning indicate the involvement of autophagy in cardioprotection against exercise-induced myocardial injury J Physiol Sci Feb 17 2020 [PMID: 32066368] (IHC, Rat)
• 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
• Dong W, Luo Y, Zhang G et al. Carbon Nanospheres Exert Antitumor Effects Associated with Downregulation of 4E-BP1 Expression on Prostate Cancer Int J Nanomedicine Aug 5 2020 [PMID: 32848387] (WB, Human)
• Nettesheim A, Dixon A, Shim MS et al. Autophagy in the Aging and Experimental Ocular Hypertensive Mouse Model Invest. Ophthalmol. Vis. Sci. Aug 3 2020 [PMID: 32797200] (WB, Mouse)
• Lindahl M, Chalazonitis A, Palm E et al. Cerebral dopamine neurotrophic factor-deficiency leads to degeneration of enteric neurons and altered brain dopamine neuronal function in mice Neurobiol Dis. Nov 26 2019 [PMID: 31783118] (IHC-WhMt, Mouse)
• Sarnyai F, SzekerczEs T, Csala M et al. BGP-15 Protects Mitochondria in Acute, Acetaminophen Overdose Induced Liver Injury Pathol. Oncol. Res. Nov 8 2019 [PMID: 31705481] (IHC-P, Mouse)
• Bryant J, Batis N, Franke AC et al. Repurposed quinacrine synergizes with cisplatin, reducing the effective dose required for treatment of head and neck squamous cell carcinoma Oncotarget Aug 27 2019 [PMID: 31497252] (WB, Human)
• Ding Y, Dvornikov AV, Ma X et al. Haploinsufficiency of mechanistic target of rapamycin ameliorates bag3 cardiomyopathy in adult zebrafish Dis Model Mech Sep 6 2019 [PMID: 31492659] (WB, Zebrafish)
• Yong T, Zhang X, Bie N et al. Tumor exosome-based nanoparticles are efficient drug carriers for chemotherapy Nat Commun Aug 23 2019 [PMID: 31444335] (WB, Human)
• Gregosa A, Vinuesa A, Todero MF et al. Periodic dietary restriction ameliorates amyloid pathology and cognitive impairment in PDAPP-J20 mice: Potential implication of glial autophagy Neurobiol. Dis. Jul 24 2019 [PMID: 31351172] (ICC/IF, IHC-Fr, Mouse)
• Liou YF, Chen PN, Chu SC et al. Thymoquinone suppresses the proliferation of renal cell carcinoma cells via reactive oxygen species-induced apoptosis and reduces cell stemness Environ. Toxicol. Jul 12 2019 [PMID: 31298468]
• Di Pardo A, Pepe G, Castaldo S et al. Stimulation of Sphingosine Kinase 1 (SPHK1) Is Beneficial in a Huntington's Disease Pre-clinical Model Front Mol Neurosci Apr 24 2019 [PMID: 31068790] (WB, Mouse)
• Morelli, MB;Amantini, C;Tomassoni, D;Nabissi, M;Arcella, A;Santoni, G; Transient Receptor Potential Mucolipin-1 Channels in Glioblastoma: Role in Patient's Survival Cancers (Basel) Apr 12 2019 [PMID: 31013784] (WB, Human)
• Zhang R, Varela M, Vallentgoed W et al. The selective autophagy receptors Optineurin and p62 are both required for zebrafish host resistance to mycobacterial infection PLoS Pathog. 2019 Feb 27 [PMID: 30818338] (WB, Zebrafish)
• Hoffmann AC, Minakaki G, Menges S et al. Extracellular aggregated alpha synuclein primarily triggers lysosomal dysfunction in neural cells prevented by trehalose. Sci Rep Jan 24 2019 [PMID: 30679445] (WB, Human)
• Minakaki G, Canneva F, Chevessier F et al. Treadmill exercise intervention improves gait and postural control in alpha-synuclein mouse models without inducing cerebral autophagy. Behav. Brain Res. Dec 29 2018 [PMID: 30599154] (WB, Mouse)
• 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)
• Hirt J, Porter K, Dixon A et al. Contribution of autophagy to ocular hypertension and neurodegeneration in the DBA/2J spontaneous glaucoma mouse model. Cell Death Discovery Jul 17 2018 [PMID: 30210817] (WB, Mouse)
• 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)
• Maier L, Kallenberger S, Jechow K et al. Unraveling mitotic protein networks by 3D multiplexed epitope drug screening bioRxiv [PMID: 30104419] (ICC/IF, Human)
• 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)
• 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)
• NAger M, SallAn MC, Visa A et al. Inhibition of WNT-CTNNB1 signaling upregulates SQSTM1 and sensitizes glioblastoma cells to autophagy blockers Autophagy. 2018 Feb 20 [PMID: 29313411] (WB, Human)
• 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)
• Agarwal AK, Tunison K, Dalal JS et al. Metabolic, Reproductive, and Neurologic Abnormalities in Agpat1-Null Mice Endocrinology. 2017 Oct 31 [PMID: 28973305] (WB, Mouse)
• 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)
• Fu Qiang, Chen Kang, Zhu Qian et al. Beta-catenin promotes intracellular bacterial killing via suppression of Pseudomonas aeruginosa-triggered macrophage autophagy. The Journal of International Medical Research 2017 Apr [PMID: 28415949] (WB, Mouse)
• 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)
• Nuschke A, Rodrigues M, Wells AW et al. Mesenchymal stem cells/multipotent stromal cells (MSCs) are glycolytic and thus glucose is a limiting factor of in vitro models of MSC starvation Stem Cell Res Ther. 2016 Nov 30 [PMID: 27906055] (WB, Mouse)
• 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)
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  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).
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  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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  Details:
  LC3 antibody used for WB in hearts from mice injected with PBS, miR-Co or miR-144 (Figure 2b).
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Reviews for LC3A Antibody (NB100-2331) (20)

Average Rating: 4.4

(Based on 20 reviews)

We have 20 reviews tested in 4 species: Human, Mouse, Mouse and Human, Other.

reviewed by: Anonymous WB Human 06/09/2020

Summary

• Application: Western Blot
• Sample Tested: THP-1 cell lysate
• Species: Human
• Lot: AB-3

Comments

• Comments: The antibody works great to detect the LC3 from THP-1 cell lysates

reviewed by: Anonymous WB Mouse 01/29/2019

Summary

• Application: Western Blot
• Sample Tested: lysed cell lines, Sample Amount: 20 ug
• Species: Mouse
• Lot: A6-NB100-2331

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