Recombinant Human LC3A GST (N-Term) Protein

SDS-Page: Recombinant Human LC3A Protein [H00084557-P01] - 12.5% SDS-PAGE Stained with Coomassie Blue.

Product Discontinued

Recombinant Human LC3A GST (N-Term) Protein Summary

• Description: Recombinant protein with GST-tag at N-terminal corresponding to the amino acids 1 - 121 of Human MAP1LC3A full-length ORF
  
  

  Source: Wheat Germ (in vitro)

  Amino Acid Sequence: MPSDRPFKQRRSFADRCKEVQQIRDQHPSKIPVIIERYKGEKQLPVLDKTKFLVPDHVNMSELVKIIRRRLQLNPTQAFFLLVNQHSMVSVSTPIADIYEQEKDEDGFLYMVYASQETFGF

• Preparation Method: in vitro wheat germ expression system
• Details of Functionality: This protein was produced in an in vitro wheat germ expression system that should preserve correct conformational folding that is necessary for biological function. While it is possible that this protein could display some level of activity, the functionality of this protein has not been explicitly measured or validated.
• Source: Wheat germ
• Protein/Peptide Type: Full Length Recombinant Protein
• Gene: MAP1LC3A
• Purity: >80% by SDS-PAGE and Coomassie blue staining

Applications/Dilutions

• Dilutions:
  • ELISA
  • Immunoaffinity Purification
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
  • Protein Array
  • SDS-Page
  • Western Blot
• Theoretical MW: 38.94 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.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: 50 mM Tris-HCl, 10 mM reduced Glutathione, pH 8.0 in the elution buffer.
• Preservative: No Preservative
• Purity: >80% by SDS-PAGE and Coomassie blue staining

Notes

This product is produced by and distributed for Abnova, a company based in Taiwan.

Alternate Names for Recombinant Human LC3A GST (N-Term) Protein

• 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. Peptides and proteins are guaranteed for 3 months from date of receipt.

Publications for LC3A Full Length Recombinant Protein (H00084557-P01)(6)

Publications using H00084557-P01

• Korschgen H, Baeken M, Schmitt D et al. Co-chaperone BAG3 directly target autophagic degradation via its LC3-interacting regions bioRxiv 2023-02-03 (WB, PLA)
• Koukourakis MI, Kalamida D, Giatromanolaki A et al. Autophagosome Proteins LC3A, LC3B and LC3C Have Distinct Subcellular Distribution Kinetics and Expression in Cancer Cell Lines. PLoS One 2015-01-01 [PMID: 26378792]
• Giatromanolaki A, Sivridis E, Mitrakas A et al. Autophagy and lysosomal related protein expression patterns in human glioblastoma. Cancer Biol Ther. 2014-01-01 [PMID: 25482944] (Human)
• Koukourakis MI, Giatromanolaki A, Bottini A et al. Prospective neoadjuvant analysis of PET imaging and mechanisms of resistance to Trastuzumab shows role of HIF1 and autophagy. Br J Cancer 2014-04-10 [PMID: 24722179] (IF/IHC, Human)
• Koukourakis MI, Giatromanolaki A, Zois CE, Sivridis E. LC3 immunostaining pitfalls. Histopathology. 2013-05-01 [PMID: 23530618]
• Martinet W, Schrijvers DM, Timmermans JP et al. Immunohistochemical analysis of macroautophagy: Recommendations and limitations. Autophagy 2012-12-14 [PMID: 23242143]

FAQs for LC3A Full Length Recombinant Protein (H00084557-P01). (Showing 1 - 1 of 1 FAQs).

1. One customer received our notification that the application of H00084557-P01 is changed when he tried to order a new one. And he got good result with the previous one. So he would like to know whether the formulation or lot has changed? If it is due to lack of validation, I think the customer would like to purchase it.
   • Abnova has recently removed these applications from their product page due to lack of validation. To my understanding these were listed based on references and Abnova is not doing Quality Control for these applications. The formulation has not changed. I see we have updated the buffer to "50 mM Tris-HCI, 10 mM reduced Glutathione, pH 8.0" because it was not listed, not because it has changed. If you customer has been using this protein successfully, I do not think there is any reason to think that it will suddenly fail.

Bioinformatics

• Gene Symbol: MAP1LC3A