Biological Strategies: Western Blot: LC3A Antibody [NBP1-19167] - Total protein from HeLa and Neuro2A cells treated with or without 50 uM chloroquine for 24 hours was separated on a 4-15% gel by SDS-PAGE, ...read more
Immunocytochemistry/ Immunofluorescence: LC3A Antibody [NBP1-19167] - LC3/MAP1 [NBP1-19167] - LC3 antibody was tested in HeLa cells with Dylight 488 (green). Cells were treated overnight with 50 uM chloroquine to induce ...read more
Immunohistochemistry-Paraffin: LC3A Antibody [NBP1-19167] - Analysis of a FFPE tissue section of mouse brain using LC3 antibody at 1:300 dilution. The signal was developed using HRP-labelled secondary antibody and DAB ...read more
Western Blot: LC3/MAP1LC3A Antibody [NBP1-19167] - Human brain lysate.
Immunohistochemistry-Paraffin: LC3A Antibody [NBP1-19167] - Analysis of a FFPE tissue section of mouse liver using LC3 antibody at 1:300 dilution. The signal was developed using HRP-labelled secondary antibody and DAB ...read more
Simple Western: LC3A Antibody [NBP1-19167] - 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.
In WB, bands are seen at approx. 14-17 kDa. In ICC/IF, autophagosome formation has been seen in HeLa cells after treatment with 50 uM chloroquine. Use in IHC-Fr reported in scientific literature (PMID: 23936035).
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. 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.
14 kDa. Disclaimer note: 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.
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 (NBP1-19167). (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.
We have received an antibody against LC3 (# NBP1-19167) and I need a precision regarding its specificity. According to the datasheet, the antibody was raised against a peptide 100% identical to LC3A and 62% to LC3B. My question is: Is LC3A used as a synonym for LC3-I and LC3B as a synonym of LC3-II?
LC3A is not a synonym for LC3-I or LC3-II. When we refer to LC3-I and -II, we actually mean LC3B-I and LC3B-II. I hope this helps, but please let me know if you have any questions.
We purchased an antibody against LC3 (NBP1-19167) from your company. According to the information found on your web site, three papers used this antibody to detect both the LC3-I and LC3-II form of the protein. The catalog number of the antibody is not mentioned in the papers. How can I be sure that it is the one used by the authors? It is important since I want to know if this antibody NBP1-19167 recognize both LC3 forms.
Our publication evaluation team is highly qualified and we found that these publications were cited under # NBP1-19167 after consulting the archived order information of the customers. As far as the detection of the two forms is concerned, any given LC3 antibody should detect LC3-I as well as LC3-II forms because the only difference between the two forms is that LC3-II carries the phosphatidylethanolamine (PE) conjugate whereas LC3-I does not have a PE conjugate. In nutshell, following translation, the unprocessed form of LC3 (pro-LC3) is proteolytically cleaved by Atg4 protease, resulting in the cytosolic form of LC3 (LC3-I) with a carboxyterminal exposed glycine. Upon induction of autophagy, the 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 the highly lipophilic PE moiety to generate LC3-II (which is recruited to autophagosomal membranes). Therefore, the presence of both bands (LC3-I & LC3-II) will depend upon the extent of autophagy in your particular samples.
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.
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.