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LC3A Antibody [CoraFluor™ 1]

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Product Details

Summary
Reactivity Hu, Mu, Rt, Bv, ZeSpecies Glossary
Applications WB, Simple Western, Flow, ICC/IF, IHC
Clonality
Polyclonal
Host
Rabbit
Conjugate
CoraFluor 1

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LC3A Antibody [CoraFluor™ 1] Summary

Description
CoraFluor(TM) 1 is a high performance terbium-based TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) or TRF (Time-Resolved Fluorescence) donor for high throughput assay development. CoraFluor(IM) 1 absorbs UV light at approximately 340 nm, and emits at approximately 490 nm, 545 nm, 585 nm and 620 nm. It is compatible with common acceptor dyes that absorb at the emission wavelengths of CoraFluor(TM) 1. CoraFluor(TM) 1 can be used for the development of robust and scalable TR-FRET binding assays such as target engagement, ternary complex, protein-protein interaction and protein quantification assays.
Immunogen
Partial recombinant protein made to an N-terminal portion of the human LC3A protein [Swiss-Prot# Q9H492].
Localization
Autophagosomes. Cytoplasm. Intracytoplasmic membrane; Lipid-anchor. Cytoplasmic vesicle autophagosome membrane; Lipid-anchor. Note= LC3-II binds to the autophagic membranes.
Marker
Autophagosome Marker
Specificity
Although specificity between this LC3A Antibody and LC3B has not been tested, this antibody was created to a peptide that has 100% identity to LC3A and 62% identity to LC3B.
Isotype
IgG
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.

Applications/Dilutions

Dilutions
  • Flow Cytometry
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin
  • Simple Western
  • Western Blot
Application Notes
Optimal dilution of this antibody should be experimentally determined.

Packaging, Storage & Formulations

Storage
Store at 4C in the dark. Do not freeze.
Buffer
PBS
Preservative
No Preservative
Purity
Immunogen affinity purified

Notes

CoraFluor (TM) is a trademark of Bio-Techne Corp. Sold for research purposes only under agreement from Massachusetts General Hospital. US patent 2022/0025254

Alternate Names for LC3A Antibody [CoraFluor™ 1]

  • Apg8
  • APG8a
  • Apg8p3
  • ATG8E
  • Autophagy-related protein LC3 A
  • Autophagy-related ubiquitin-like modifier LC3 A
  • LC3
  • LC3A
  • MAP1 light chain 3-like protein 1
  • MAP1A/1B light chain 3 A
  • MAP1A/MAP1B LC3 A
  • MAP1A/MAP1B light chain 3 A
  • MAP1ALC3
  • MAP1BLC3
  • MAP1LC3A
  • Microtubule-associated protein 1 light chain 3 alpha
  • microtubule-associated proteins 1A/1B light chain 3
  • microtubule-associated proteins 1A/1B light chain 3A
  • MLP3A

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.

References

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

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.

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Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for LC3A Antibody (NBP1-19167CL1). (Showing 1 - 1 of 1 FAQ).

  1. 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.

Secondary Antibodies

 

Isotype Controls

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Bioinformatics

Gene Symbol MAP1LC3A