Anti-Atg12 produced in rabbit, IgG fraction of antiserum

Product Number A8731

Product Description to Atg10, another E2-like enzyme, and conjugated to Anti-Atg12 is produced in rabbit using as immunogen a Atg5.9 This Atg12-Atg5 conjugate is formed via an synthetic peptide corresponding to amino acids 27-39 of isopeptide bond between the C-terminal glycine of Atg12 mouse Atg12 (GeneID: 67526), conjugated to KLH. The and Lys130 of Atg5.7,8 The Atg12-Atg5 conjugates further corresponding sequence differs by one amino acid in to form a ~800 kDa complex with the multimeric human. Whole serum is purified using protein A protein Atg16L. The Atg12-Atg5.Atg16 multimeric immobilized on agarose to provide the IgG fraction of complex is essential for autophagosome formation.10 antiserum. Reagent Anti-Atg12 recognizes mouse Atg12 and Atg12-Atg5. Supplied as a solution in 0.01 M phosphate buffered Applications include the detection of mouse Atg12 (~ 20 saline, pH 7.4, containing 15 mM sodium azide as a kDa) and Atg12-Atg5 complex by immunoblotting and preservative. immunoprecipitation. Detection of the Atg12 and Atg12-Atg5 complex bands by immunoblotting are Precautions and Disclaimer specifically inhibited by the immunizing peptide. This product is for R&D use only, not for drug, household, or other uses. Please consult the Material Macroautophagy, usually referred to as , is a Safety Data Sheet for information regarding hazards major pathway for bulk degradation of cytoplasmic and safe handling practices. constituents and organelles. In this process, portions of the cytoplasm are sequestered into double membrane Storage/Stability vesicles, the autophagosomes, and subsequently For continuous use, store at 2-8 °C for up to one month. delivered to the lysosome for degradation and For extended storage, freeze in working aliquots. recycling.1,2 Although autophagy is a constitutive cellular Repeated freezing and thawing, or storage in “frost- event, it is enhanced under certain conditions such as free” freezers, is not recommended. If slight turbidity starvation, hormonal stimulation and drug treatments.3 occurs upon prolonged storage, clarify the solution by Autophagy is required for normal turnover of cellular centrifugation before use. Working dilutions should be components during starvation. It plays an essential role in discarded if not used within 12 hours. cellular differentiation, cell death and aging. Defective autophagy may contribute to certain human diseases Product Profile such as cancer, neurodegenerative diseases, muscular Immunoblotting: a working dilution of 1:250-1:500 is disorders and pathogen infections.4,5 Autophagy is an recommended using a whole extract of HEK-293T cells evolutionarily conserved pathway seen in all eukaryotic expressing mouse Atg12. cells.1 At least 16 ATG required for autophagosome formation were identified in yeast by Immunoprecipitation: a working amount of 5-10 mL is genetic screens. For many of these genes, related recommended using a whole extract of HEK-293T cells 6 homologs have been identified in mammals. co-expressing mouse Atg12 and Atg5-EGFP . Atg12 is a -like modifier essential for 7,8 autophagy. Two ubiquitin-like conjugation systems are Note: In order to obtain the best results using various involved in autophagosome formation: Atg12 and Atg8 techniques and preparations, we recommend conjugation systems. The ubiquitin-like proteins Atg8 and determining optimal working dilutions by titration. Atg12 are activated by Atg7 (an E1-like enzyme essential for both conjugation systems). Atg8 is then transferred to References the E2-like enzyme Atg3 and conjugated to 1. Klionsky, D.J., and Emr, S.D., Science, 290, 1717- phosphatidylethanolamine, whereas Atg12 is transferred 1721 (2000). 2. Kuma, A., et al., Nature, 432, 1032-1036 (2004). 3. Kabeya, Y., et al., EMBO J., 19, 5720-5728 (2000). 8. Hanada, T., and Yoshinori, O., Autophagy, 1, 110- 4. Reggiori, F., and Klionsky, D.J., Eukaryotic Cell, 1, 118 (2005). 11-21 (2002). 9. Komatsu, M., et al., J. Biol. Chem., 276, 9846- 5. Shintani, T., and Klionsky, D.J., Science, 306, 990- 9854 (2001). 995 (2004). 10. Mizushima, N., et al., J. Cell Sci., 116, 1679-1688 6. Klionsky, D.J., et al., Develop. Cell, 5, 539-545 (2003). (2003). 7. Mizushima, N., et al., J. Biol. Chem., 273, 33889- ST,BKR,PHC 06/08-1 33892 (1998).

Sigma brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip.