Immunocytochemistry/ Immunofluorescence: LC3A Antibody [NBP1-78964] - IF Confocal analysis of A549 cells using LC3I antibody (NBP1-78964, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary ...read more
Western Blot: LC3A Antibody [NBP1-78964] - Analysis of LC3I in Neuro2A cell lysate.
Immunocytochemistry/ Immunofluorescence: LC3A Antibody [NBP1-78964] - LC3I antibody was tested in Neuro2a cells with FITC (green). Nuclei were counterstained with DAPI (blue).
Immunohistochemistry: LC3A Antibody [NBP1-78964] - Analysis of LC3I in mouse testis using DAB with hematoxylin counterstain.
This LC3I antibody is useful for Western Blot, Immunocytochemistry/Immunofluorescence, and IHC-paraffin embedded sections. In Western Blot, a band is seen ~15kDa representing LC3I. In ICC/IF, observed staining showed inactivated LC3 throughout the cytoplasm of Neuro2a cells. In IHC-P, staining was observed in the cytoplasm of mouse testes tissue. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
Autophagy is a process of intracellular bulk degradation in which cytoplasmic components, including organelles, are sequestered within double-membrane vesicles that deliver the contents to the lysosome/vacuole for degradation. During macroautophagy, the sequestering vesicles, termed autophagosomes, fuse with the lysosome or vacuole resulting in the delivery of an inner vesicle (autophagic body) into the lumen of the degradative compartment. There are 16 proteins participating in the autophagy pathway in human. The autophagy protein LC3, a mammalian homologue of Atg8, was originally identified as microtubule-associated protein 1 light chain 3. It is a component of both the MAP1A and MAP1B microtubule-binding domains and the heavy-chain independent regulation of LC3 expression might modify MAP1 microtubule-binding activity during development. LC3 is the only known mammalian protein identified that stably associates with the autophagosome membranes. LC3-I is cytosolic and LC3-II is membrane bound and enriched in the autophagic vacuole fraction. The detection of LC3-I to LC3-II conversion is a useful and sensitive marker for distinguishing autophagy in mammalian cells.
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.
IF Confocal analysis of A549 cells using LC3I antibody (NBP1-78964, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
FAQs for LC3A Antibody (NBP1-78964). (Showing 1 - 1 of 1 FAQ).
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.
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