ELISA, Immunogen, SDS-PAGE, WB, Bioactivity testing in progress
Species
Homo sapiens (Human)
Nature
Recombinant
Endotoxin level
Please contact with the lab for this information.
Purity
>90% as determined by SDS-PAGE.
Predicted molecular weight
47.54 kDa
Form
Lyophilized
Storage buffer
Lyophilized from a solution in PBS pH 7.4, 0.02% NLS, 1 mM EDTA, 4% Trehalose, 1% Mannitol.
Please refer to the specific buffer information in the hardcopy of datasheet or the lot-specific COA.
Reconstitution
Reconstitute in sterile water for a stock solution. Adjust the volume of sterile water so that, after reconstitution, the final stock concentration matches the value indicated on the product label or datasheet.
RECONSTITUTION PROTOCOL
1. Allow the product to be reconstituted and the reconstitution diluent to reach room temperature.
2. Tap or briefly centrifuge the product vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.
3. Use the diluent and stock concentration recommended in the product datasheet. If preparing a stock concentration higher than recommended, contact Technical Service.
4. After adding the diluent, re-cap the vial and invert gently by hand or place on a slow rocking platform to allow the reconstitution diluent to coat all the surfaces inside the vial. Note: Do not mix by vortexing or by pipetting the material up and down.
5. Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.
6. Store the reconstituted material in single use aliquots in polypropylene or siliconized tubes. Aliquots of 10 μL or larger are recommended.
Shipping
In general, proteins are provided as lyophilized powder/frozen liquid. They are shipped out with dry ice/blue ice unless customers require otherwise.
Stability and Storage
Use a manual defrost freezer and avoid repeated freeze thaw cycles. Store at 2 to 8°C for frequent use. Store at -20 to -80°C for twelve months from the date of receipt.
Cysteine protease ATG4A is a ~45 kDa protein. Cysteine protease that plays a key role in autophagy by mediating both proteolytic activation and delipidation of ATG8 family proteins. The protease activity is required for proteolytic activation of ATG8 family proteins: cleaves the C-terminal amino acid of ATG8 proteins to reveal a C-terminal glycine. Exposure of the glycine at the C-terminus is essential for ATG8 proteins conjugation to phosphatidylethanolamine (PE) and insertion to membranes, which is necessary for autophagy. Preferred substrate is GABARAPL2 followed by MAP1LC3A and GABARAP. Protease activity is also required to counteract formation of high-molecular weight conjugates of ATG8 proteins (ATG8ylation): acts as a deubiquitinating-like enzyme that removes ATG8 conjugated to other proteins, such as ATG3.
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2. Kabeya, Y. et al. (2004) Journal of cell science 117, 2805-12. PMID: 15169837
3. Scherz-Shouval, R. et al. (2007) The EMBO journal 26, 1749-60. PMID: 17347651
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6. Li, M. et al. (2012) Autophagy 8, 401-12. PMID: 22302004
7. Nguyen, N. et al. (2020) The Journal of biological chemistry 295, 13584-13600. PMID: 32732290
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