Recombinant Human HA-GATE-16 AMC Protein, CF

Discontinued Product

UL-445 has been discontinued.
View all GATE-16 products.
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Recombinant Human HA-GATE-16 AMC Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Activity
Recombinant Human HA-GATE-16 AMC is a fluorogenic substrate for some ATG-specific isopeptidases.  Release of AMC fluorescence can be monitored with an excitation wavelength of 345 nM and an emission wavelength of 445 nM. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human HA-GATE-16 AMC concentration of 0.1-1 μM.
Source
E. coli-derived human GATE-16 protein
Contains an N-terminal HA (YPYDVPDYA) tag
Accession #
Predicted Molecular Mass

14 kDa

Product Datasheets

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UL-445

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

UL-445

Formulation

X mg/ml (X µM) in 50 mM MOPS pH 6.5, 100 mM NaCl

Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
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Background: GATE-16

Golgi-associated ATPase Enhancer of 16 kDa (GATE-16), also known as GABARAPL2, is a 117 amino acid (aa) polypeptide and a member of the Autophagy-related 8 (ATG8) family of proteins (1). GATE-16 has 100% aa sequence identity with its mouse and rat orthologs, and is orthologous to the yeast ATG8. ATG8 family members show structural similarity with Ubiquitin, but lack aa sequence similarity. GATE-16 is best known for its role in autophagy (2,3). GATE-16 covalently attaches to phosphatidylethanolamine (PE) the phagophore (autophagosome precursor) membrane using a Ubiquitin-like conjugation system that includes Ubiquitin-activating (E1)-, Ubiquitin-conjugating (E2)-, and Ubiquitin Ligase (E3)-like enzymes. Here it is involved in the later stages of autophagosome formation (4,5). It may also be involved in cargo recruitment to autophagosomes (1).

 

This is a fluorogenic substrate for ATG8-specific C-terminal hydrolases such as ATG4B (Catalog # E-400) based on the C-terminal derivatization of ATG8 with 7-amido-4-methylcoumarin (AMC). ATG8-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. Wang, H. et al. (1999) Nature 397:69.
  3. Leil, T.A. et al. (2004) J. Neurosci. 24:11429.
  4. Weidberg, H. et al. (2010) EMBO J. 29:1792.
  5. Weidberg, H. et al. (2011) Ann. Rev. Biochem. 80:125.
Long Name
Golgi-associated ATPase Enhancer of 16 kDa
Entrez Gene IDs
11345 (Human); 100042211 (Mouse); 64670 (Rat)
Alternate Names
Apg8p2; ATG8; ATG8C; FLC3A; GABARAPL2; gamma-aminobutyric acid receptor-associated protein-like 2; Ganglioside expression factor 2; GATE16; GATE-16; GEF2; GEF-2; GEF2GEF-2; General Protein Transport Factor P16; Golgi-Associated ATPase Enhancer Of 16 KDa; MAP1 light chain 3 related protein; MAP1 light chain 3-related protein

Citation for Recombinant Human HA-GATE-16 AMC Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Cholesterol impairs autophagy-mediated clearance of amyloid beta while promoting its secretion
    Authors: E Barbero-Ca, V Roca-Aguje, I Bartolessi, C de Dios, JC Fernández-, M Marí, A Morales, T Hartmann, A Colell
    Autophagy, 2018-06-04;0(0):1-26.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Bioassay

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