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 ...read more
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 ...read more
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 ...read more
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 ...read more
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 ...read more
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 ...read more
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.
Hu, Mu, Rt, Am, Ca, Fi, Pl, Ze, BvSpecies Glossary
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).
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.
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
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.
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.
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