Note: Mouseover a species abbreviation on the product page to display the fullname. 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. Product Details Summary Reactivity Hu, Mu, Rt, Am, Ca, Fi, Pl, Ze, BvSpecies Glossary Applications WB, Simple Western, ChIP, 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 Order Details SizePrice 0.025 mlNB100-2331SSContact YourDistributor In Stock: 6-8 Business Days 0.1 mlNB100-2331Contact YourDistributor In Stock: 6-8 Business Days 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 Clonality Polyclonal Host Rabbit Gene MAP1LC3A Purity Immunogen affinity purified Innovator\'s Reward Test in a species/application not listed above to receive a full credit towards a future purchase.Learn about the Innovator\'s Reward Western Blot Simple Western 1:50Chromatin Immunoprecipitation ELISA Flow Cytometry Immunoblotting Immunocytochemistry/Immunofluorescence 1:100-1:300Immunohistochemistry 1:200-1:400Immunohistochemistry-Frozen Immunohistochemistry-Paraffin 1:200-1:400Immunoprecipitation 20 ug / 500 ug of lysateSouthern 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. Use in Chromatin Immunoprecipitation reported in scientific literature (PMID:33035707). Use in Rat reported in scientific literature (PMID:33678798). Use in Amphibian reported in scientific literature (PMID:29777142). Packaging, Storage & Formulations Storage Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles. Buffer Preservative 0.02% Sodium Azide Concentration 1.0 mg/ml Purity Immunogen affinity purified Alternate Names for LC3A Antibody Apg8APG8aApg8p3ATG8EAutophagy-related protein LC3 AAutophagy-related ubiquitin-like modifier LC3 ALC3LC3AMAP1 light chain 3-like protein 1MAP1A/1B light chain 3 AMAP1A/MAP1B LC3 AMAP1A/MAP1B light chain 3 AMAP1ALC3MAP1BLC3MAP1LC3AMicrotubule-associated protein 1 light chain 3 alphamicrotubule-associated proteins 1A/1B light chain 3microtubule-associated proteins 1A/1B light chain 3AMLP3A 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. References1. 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-2011-12-7-2262. 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.01376753. Weidberg, H., Shvets, E., & Elazar, Z. (2011). Biogenesis and cargo selectivity of autophagosomes. Annual review of biochemistry. https://doi.org/10.1146/annurev-biochem-052709-0945524. 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.2004.05.0095. 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. Customers Who Viewed This Item Also Viewed... (248 Publications) Species: Hu, Mu, Rt, Bv, Ha, RbApplications: WB, ELISA, ICC/IF, IHC, IHC-Fr, IHC-P, IP(119 Publications) Species: Hu, Mu, RtApplications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KD(2 Publications) Species: HuApplications: WB, ELISA, ICC/IF, IHC, IHC-P, S-ELISA, KD(2 Publications) Species: HuApplications: WB, ICC/IF, IHC, IHC-P(2 Publications) Species: Hu, Bv, MaApplications: WB, ICC/IF, IHC, IHC-P, IP(9 Publications) Species: Hu, RtApplications: WB, ELISA, IHC, IHC-P(220 Publications) Species: Hu, Mu, Rt, Po, Al, Bv, Dr, Fi, Gp, Pm, Xp, ZeApplications: WB, Simple Western, EM, ELISA, Flow, IB, ICC/IF, IHC, IHC-P, IP, PLA, RIA, KD, KO(158 Publications) Species: Hu, Mu, Rt, Po, Ch, ChHa, MaApplications: WB, Simple Western, EM, Flow, HEStain, IB, ICC/IF, IHC, IHC-Fr, IHC-P, CyTOF-ready, IHC-FrFl, KD, KO(25 Publications) Species: Hu, Mu, RtApplications: WB, Simple Western, IHC, CyTOF-ready, ICFlow(4 Publications) Species: Hu, MuApplications: WB, Simple Western, IHC, CyTOF-ready, ICC, ICFlow(11 Publications) Species: Hu, Mu, Rt, CaApplications: WB, IHC, IHC-P, IP(26 Publications) Species: Hu, Mu, RtApplications: WB, ICC/IF, IHC, IHC-P(33 Publications) Species: Hu, Mu, Rt, Ye, Xp(-)Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, CyTOF-ready, Flow-IC Publications for LC3A Antibody (NB100-2331)(265) We have publications tested in 8 confirmed species: Human, Mouse, Rat, Amphibian, Canine, Fish, Plant, Zebrafish.We have publications tested in 12 applications: ChIP, ELISA, FLOW, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IHC-WhMt, IP, WB, WB,IP. Submit a Publication 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 We have 22 reviews tested in 5 species: Human, Mouse, Rat, Human and Mouse, Other. Submit a Review Reviews using NB100-2331: Filter by Applications ICC (1) WB (18) IF (1) IHC-P (1) IHC (1)All Applications Summary ApplicationImmunocytochemistrySample TestedHuman microglia,Human microglia cell lysateSpeciesHumanLotAK Summary ApplicationWestern BlotSample TestedTHP-1 cell lysateSpeciesHumanLotAB-3Comments CommentsThe antibody works great to detect the LC3 from THP-1 cell lysates Summary ApplicationWestern BlotSample Testedlysed cell lines, Sample Amount: 20 ugSpeciesMouseLotA6-NB100-2331 Summary ApplicationWestern BlotSample Testedmouse hepatocytes and HepG2 cellsSpeciesHuman and MouseLotAC-1 Summary ApplicationWestern BlotSample TestedMouse macrophage cell line RAW 264.7SpeciesMouseLotH6-NB100-2331 Summary ApplicationWestern BlotSample TestedMouse Neuro2A, Mouse Cortical Neurons whole cell lysateSpeciesMouseLotR-4Blocking Blocking Details10% non-fat dry milk in TBS-T (0.05% Tween) for 1hr at room temperaturePrimary Anitbody Dilution Ratio1:1000Secondary Antibody Secondary DescriptionGoat-anti rabbit HRPSecondary Concentration1:5000 in 3% milk in TBS-T (0.05%)Details Detection NotesClarity ECL, Biorad Chemidoc imager, exposed for 1 min, Neg control= untreated cells, Pos Control= rapamycinComments CommentsThis antibody works very well. Very strong clear signal. ApplicationWestern BlotSample Testedhuman glioblastomaSpeciesHumanBlocking Blocking DetailsLi-Cor blockin buffferPrimary Anitbody Dilution Ratio2 ug/ml, overnight, room temperature, TBST bufferSecondary Antibody Secondary Descriptionanti-IgG secondary antibodies, Conjugation: IRDye® 800CWSecondary Manufacturer Cat#Li-CorSecondary Concentrationper vendor recomendationsDetails Detection NotesOdyssey Li-CorComments CommentsPlease site our paper:Tamoxifen improves cytopathic effect of oncolytic adenovirus in primary glioblastoma cells mediated through autophagy.\" just accepted for Oncotarget. ApplicationWestern BlotSample TestedSee PMID 22892563SpeciesMouseBlocking Blocking DetailsSee PMID 22892563Details Detection NotesSee PMID 22892563Comments CommentsPublished in PMID: 22892563 Summary ApplicationImmunofluorescenceSample TestedNIH3T3, HeLa, HEK293T, N2a neuroblastoma, Q7 striatal, Sy5Y neuroblastomaSpeciesMouseBlocking Blocking Details1% BSA + 1%milkPrimary Anitbody Dilution Ratio1:100 for IFSecondary Antibody Secondary DescriptionCy5 conjuagated anti-rabbitSecondary Concentration1:500Details Detection NotesImages taken by Zeiss Axiovert microscopeFixation DetailsPFA Fixed, 0.1% Tween20 permeabilized, PBS, 1hr primary incubation at RTWash DescriptionPBS 3x washes, 2 mins each Summary ApplicationImmunohistochemistry-ParaffinLotS-1CommentsThese tissue samples were fixed with zinc buffered formalin. Summary ApplicationWestern BlotSample Testedhuman tumor cellsSpeciesHumanLotq-1Blocking Blocking Details#917-40000, LI-CORPrimary Anitbody Dilution Ratioin agreement with vendor recommendationSecondary Antibody Secondary Descriptiongoat-anti-rabbitSecondary Manufacturer Cat#Li-CORDetails Detection NotesOdyssey Imaging System (Model #1866, Li-COR Biosciences Summary ApplicationWestern BlotSpeciesHumanPub Med ID24735980FileView PDF Summary ApplicationWestern BlotSpeciesHumanLotS-2Special ApplicationsGives slighter LC3-I band compared to LC3II. Stronger binding to LC3II. High level of background at first usage in BSA solution.Pub Med ID24358205FileView PDF Summary ApplicationImmunohistochemistrySample TestedRenal tissue (paraffin block)SpeciesMouseLotNCommentsAntibody 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 DetailsProtein block, Dako, 20 min RTPrimary Anitbody Dilution Ratio1:10,000, 4dC, ON, 0.1%BSA plus0.05% skim milkSecondary Antibody Secondary DescriptionGoat anti-rabbit-HRPSecondary Manufacturer Cat#DAKO, #K4011Secondary ConcentrationDAKO kitDetails Detection NotesDAB, 5 minFixation Details10% NBF, 10mM Sodium Citrate buffer pH6.0Wash DescriptionTBS-T, 5 min X 4Comments CommentsAntibody is good for immunohistochemistry. But, since this is polyclonal, each lot has to be tested for concentration of primary antibody to get desired results. Summary ApplicationWestern BlotSample TestedHumanSpeciesHumanCommentsGood antibody!Blocking Blocking DetailsBlocking Buffer: 10% non-fat milkPrimary Anitbody Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnightSecondary Antibody Secondary DescriptionSecondary Ab: rabbitDetails Detection NotesDetection Method: HRPComments CommentsGood antibody! ApplicationWestern BlotSample TestedHumanSpeciesHumanBlocking Blocking DetailsBlocking Buffer: 5% MilkPrimary Anitbody Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees CelsiusSecondary Antibody Secondary DescriptionSecondary Ab Dilution Ratio: 1:5000Details Detection NotesDetection Method: ECL-HRP ApplicationWestern BlotSample TestedHumanSpeciesHumanBlocking Blocking DetailsBlocking Buffer: 2% Skim MilkPrimary Anitbody Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees CelsiusSecondary Antibody Secondary DescriptionSecondary Ab: Goat anti-rabbit HRPDetails Detection NotesDetection Method: ECL Summary ApplicationWestern BlotSample TestedMouseSpeciesMouseCommentsThe antibody worked well!Blocking Blocking DetailsBlocking Buffer: 5% MilkPrimary Anitbody Dilution RatioPrimary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: 16 hoursSecondary Antibody Secondary DescriptionSecondary Ab: anti-mouse HRP antibodyDetails Detection NotesDetection Method: ECLComments CommentsThe antibody worked well! Summary ApplicationWestern BlotSample Tested3T3-L1 adipocyte, Sample Amount: 30ugSpeciesMouseLotH2CommentsI used in for 3T3-L1 mouse adipocyte and it is working great.Blocking Blocking DetailsBlocking Buffer: 5% Milk in PBST, Blocking Time: 1 hour, Blocking Temp: Room temperaturePrimary Anitbody Dilution RatioDilution Ratio: 1:1000, Incubation Dilution Buffer: PBST, Incubation Time: 2 hours, Incubation Temp: room temperatureSecondary Antibody Secondary DescriptionSecondary Ab: Rabbit, Secondary Ab Dilution Ratio: 1:3000Secondary Manufacturer Cat#Santa Cruz sc-2077Details Detection NotesDetection Method: ECL - perkin elmer, Exposure Time: 1 minute, Positive Control: Actin, Specific Bands: 15, 18 kDaComments CommentsI used in for 3T3-L1 mouse adipocyte and it is working great. Summary ApplicationWestern BlotSample Testedlysed cell lines, Sample Amount: 20 ugSpeciesOtherLotJBlocking Blocking DetailsBlocking Buffer: TBS 0.1% Tween20 5% NFDM, Blocking Time: 1 hour, Blocking Temp: Room temperaturePrimary Anitbody Dilution RatioDilution Concentration: 0.002, Incubation Dilution Buffer: TBS 0.1% Tween20 5% NFDM, Incubation Time: 1 hr, Incubation Temp: RTSecondary Antibody Secondary DescriptionSecondary Ab: Goat anti-Rabbit HRP, Secondary Ab Dilution Ratio: 1/4000Details Detection NotesDetection Method: Super Signal West Dura, Exposure Time: 1 minute, Specific Bands: 19 and 17kDa doublet Summary ApplicationWestern BlotSample TestedHuman cancer cells, Sample Amount: 80ugSpeciesHumanLotSSBlocking Blocking DetailsBlocking Buffer: 5% Milk in PBST, Blocking Time: 30 minutes, Blocking Temp: Room temperaturePrimary Anitbody Dilution RatioPrimary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees CelsiusDetails Detection NotesDetection Method: Licor, Exposure Time: 2 minutes, Specific Bands: 17 and 19 kDa View specific protocols for LC3A Antibody (NB100-2331): Western Blot protocol specific for LC3 Antibody (NB100-2331)Immunohistochemistry-Paraffin protocol for LC3A Antibody (NB100-2331)Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars. Western BlotImmunoprecipitationImmunohistochemistry-FrozenImmunohistochemistry-ParaffinELISAImmunocytochemistry/ImmunofluorescenceFlow CytometryChromatin ImmunoprecipitationView all Protocols, Troubleshooting, Illustrated assays and Webinars. Video Protocols WB Video Protocol ChIP Video Protocol ChIP Webinar ICC/IF Video Protocol FAQs for LC3A Antibody (NB100-2331). (Showing 1 - 3 of 3 FAQs). 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. 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. 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. The LC3 A, B, C’s and 1, 2, 3’s By Christina Towers, PhD Autophagy is a catabolic process used to breakdown and recycle damaged proteins and organelles. It is a multistep process that, in its simplest form, consists of 4 steps: initiation, phago...Read full blog post. Losing memory: Toxicity from mutant APP and amyloid beta explain the hippocampal neuronal damage in Alzheimer\'s disease By Jamshed Arslan Pharm.D. Alzheimer\'s disease (AD) is an irreversible brain disorder that destroys memory and thinking skills. The telltale signs of AD brains are extracellular deposits of amy...Read full blog post. Nuclear LC3: Why is it there and what is it doing? By Christina Towers, PhD. Cells use the complex process of autophagy to degrade and recycle cytoplasmic material.  There are over 20 proteins that have been implicated in this process and appropriately named core...Read full blog post. Why LC3B Antibodies Make Ideal Autophagosomes Membrane Markers The human form of microtubule-associated protein light chain 3 (LC3) is expressed as 3 splice variants;LC3A,LC3BandLC3C(He et al., 2003). LC3B is a subunit of theMAP1AandMAP1Bmicrotubule-binding pr...Read full blog post. The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

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