|Application ||IF, WB, IHC-P, E|
|Calculated MW||52922 Da|
|Antigen Region||441-470 aa|
|Other Names||Cysteine protease ATG4D, 3422-, AUT-like 4 cysteine endopeptidase, Autophagin-4, Autophagy-related cysteine endopeptidase 4, Autophagy-related protein 4 homolog D, Cysteine protease ATG4D, mitochondrial, ATG4D, APG4D, AUTL4|
|Target/Specificity||This ATG4D antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 441-470 amino acids from the C-terminal region of human ATG4D.|
|Format||Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is prepared by Saturated Ammonium Sulfate (SAS) precipitation followed by dialysis against PBS.|
|Storage||Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.|
|Precautions||ATG4D Antibody (C-term) is for research use only and not for use in diagnostic or therapeutic procedures.|
|Function||Cysteine protease ATG4D: Cysteine protease required for the cytoplasm to vacuole transport (Cvt) and autophagy. Cleaves the C-terminal amino acid of ATG8 family proteins MAP1LC3 and GABARAPL2, 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. Has also an activity of delipidating enzyme for the PE-conjugated forms.|
|Cellular Location||Cysteine protease ATG4D: Cytoplasm|
|Tissue Location||Mainly expressed in skeletal muscle and, to a lower extent, in testis.|
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Provided below are standard protocols that you may find useful for product applications.
Macroautophagy is the major inducible pathway for the general turnover of cytoplasmic constituents in eukaryotic cells, it is also responsible for the degradation of active cytoplasmic enzymes and organelles during nutrient starvation. Macroautophagy involves the formation of double-membrane bound autophagosomes which enclose the cytoplasmic constituent targeted for degradation in a membrane bound structure, which then fuse with the lysosome (or vacuole) releasing a single-membrane bound autophagic bodies which are then degraded within the lysosome (or vacuole). APG4 is a cysteine protease required for autophagy, which cleaves the C-terminal part of either MAP1LC3, GABARAPL2 or GABARAP, allowing the liberation of form I. A subpopulation of form I is subsequently converted to a smaller form (form II). Form II, with a revealed C-terminal glycine, is considered to be the phosphatidylethanolamine (PE)-conjugated form, and has the capacity for the binding to autophagosomes.
References for protein:
1.Baehrecke EH. Nat Rev Mol Cell Biol. 6(6):505-10. (2005)
2.Lum JJ, et al. Nat Rev Mol Cell Biol. 6(6):439-48. (2005)
3.Greenberg JT. Dev Cell. 8(6):799-801. (2005)
4.Levine B. Cell. 120(2):159-62. (2005)
5.Shintani T and Klionsky DJ. Science. 306(5698):990-5. (2004)
References for U251 cell line:
1. Westermark B.; Pontén J.; Hugosson R. (1973).” Determinants for the establishment of permanent tissue culture lines from human gliomas”. Acta Pathol Microbiol Scand A. 81:791-805. [PMID: 4359449].
2. Pontén, J.,Westermark B. (1978).” Properties of Human Malignant Glioma Cells in Vitro”. Medical Biology 56: 184-193.[PMID: 359950].
3. Geng Y.;Kohli L.; Klocke B.J.; Roth K.A.(2010). “Chloroquine-induced autophagic vacuole accumulation and cell death in glioma cells is p53 independent”. Neuro Oncol. 12(5): 473–481.[ PMID: 20406898].
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