Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Bovine, Zebrafish

Cited:

Human, Mouse

Applications

Validated:

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Simple Western, Western Blot

Cited:

IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

1.0 mg/ml

View all LC3A Antibodies »

Product Specifications

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.

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.

Marker

Autophagosome Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

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.

Scientific Data Images for LC3A Antibody

Western Blot: LC3A Antibody [NBP1-19167]

Western Blot: LC3A Antibody [NBP1-19167]

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, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-LC3A in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the detection LC3 II upon chloroquine treatment.
Immunocytochemistry/ Immunofluorescence: LC3A Antibody [NBP1-19167]

Immunocytochemistry/ Immunofluorescence: LC3A Antibody [NBP1-19167]

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 autophagosome formation. Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
Immunohistochemistry-Paraffin: LC3A Antibody [NBP1-19167]

Immunohistochemistry-Paraffin: LC3A Antibody [NBP1-19167]

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 reagent, and the sections/nuclei were further counterstained with hematoxylin. Note the diffused cytoplasmic staining of LC3 in all of the cells with highest positivity in various neurons.
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Applications for LC3A Antibody

Application
Recommended Usage

Flow Cytometry

1:100

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:100 - 1:400

Immunohistochemistry-Frozen

reported in scientific literature (PMID 23936035)

Immunohistochemistry-Paraffin

1:100 - 1:400

Simple Western

1:40

Western Blot

1:2500
Application Notes
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.

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.

Reviewed Applications

Read 2 reviews rated 4.5 using NBP1-19167 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: LC3A

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

Long Name

Microtubule-associated Protein 1 Light Chain 3 alpha

Alternate Names

Apg8, APG8a, Apg8p3, ATG8E, LC3, MAP1ALC3, MAP1LC3A, MLP3A

Entrez Gene IDs

84557 (Human); 66734 (Mouse); 64862 (Rat)

Gene Symbol

MAP1LC3A

UniProt

Q9H492

Additional LC3A Products

Product Documents for LC3A Antibody

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Product Specific Notices for LC3A Antibody

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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