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.
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.
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.
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