Recombinant Human HA-LC3A AMC Protein, CF

R&D Systems | Catalog # UL-450

R&D Systems
Discontinued Product
UL-450 has been discontinued. View all LC3A products.

Key Product Details

Source

E. coli

Accession Number

Conjugate

7-Amino-4-Methyl Coumarin

Applications

Enzyme Activity
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Product Specifications

Source

E. coli-derived human LC3A protein

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Activity

Recombinant Human HA-LC3/MAP1LC3A/Apg8p3 AMC is a fluorogenic substrate for some Apg-specific isopeptidases. Release of AMC fluorescence can be monitored with an excitation wavelength of 380 nM and an emission wavelength of 460 nM. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human HA-LC3/MAP1LC3A/Apg8p3 AMC concentration of 0.1-1 μM.

Formulation, Preparation, and Storage

UL-450
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Store the unopened product at -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: LC3A

Human Microtubule-associated Protein (MAP) Light Chain 3 (LC3) A is a121 amino acid (aa) protein with a predicted molecular weight of 14 kDa. It is a member of the LC3 subfamily of Autophagy-related 8 (Atg8) proteins (1). The LC3 subfamily also includes LC3B andLC3C. LC3 exhibits 100% aa sequence identity with its mouse and rat orthologs, and is orthologous to the yeast autophagy-related protein Atg8. Atg8 family members show structural similarity with Ubiquitin, but lack aa sequence similarity. LC3 was originally described as part is part of a complex that includes heavy and light chains comprising the MAP1 family of microtubule regulatory proteins (3). However, LC3 has gained attention for MAP1-independent functions in autophagy. LC3 utilizes a ubiquitin-like conjugation system that includes E1-, E2-, and E3-like enzymes to covalently attach phosphatidylethanolamine (PE) to its C-terminus, incorporating it into the phagophore membrane during the early stages of autophagasome formation (4). Recruitment of LC3 to the phagophore may promote membrane elongation (4,5). It may also be involved in cargo recruitment to autophagosomes (1). LC3 is often used as a marker of autophagy.

 

Fluorogenic substrate for Apg8-specific C-terminal hydrolases such as Apg4b (Catalog # E-400) based on the C-terminal derivatization of Apg8 with 7-amido-4-methylcoumarin (AMC). Apg8-AMC is useful for studying such enzyme activities when detection sensitivity or continuous monitoring of activity is essential. NOTE: this protein has an N-terminal HA tag.

References

  1. Shpilka, T. et al. (2011) Genome Biol. 12:226.
  2. He, H. et al. (2003) J. Biol. Chem. 278:29278.
  3. Kuznetsov, S.A. & V.I. Gelfand (1987) FEBS Let. 212:145.
  4. Weidberg, H. et al. (2011) Ann Rev. Biochem. 80:125.
  5. Weidberg, H. et al. (2010) EMBO J. 29:1792.

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); 362245 (Rat); 852200 (Yeast)

Gene Symbol

MAP1LC3A

Additional LC3A Products

Product Documents for Recombinant Human HA-LC3A AMC Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human HA-LC3A AMC Protein, CF

For research use only

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