Xenophagy Antibody Sampler Kit #56175
Product Information
Kit Usage Information
Protocols
- 5105: Western Blotting, Immunoprecipitation (Agarose)
- 5483: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
- 7074: Western Blotting
- 8089: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence*
- 28405: Western Blotting
- 38066: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
- 43566: Western Blotting, Immunofluorescence*
- 45511: Western Blotting
- 60732: Western Blotting
- 89572: Western Blotting, Immunoprecipitation (Magnetic), Immunohistochemistry (Paraffin), Immunofluorescence, Immunofluorescence
Product Description
Specificity / Sensitivity
Each antibody in the Xenophagy Antibody Sampler Kit detects endogenous levels of its target protein. Galectin-3/LGALS3 (E7B6R) Rabbit mAb recognizes mouse Galectin-3/LGALS3 protein and is also reactive with human Galectin-3/LGALS3; however, this antibody is not suggested for immunohistochemical analysis of human tissues. Atg16L1 (D6D5) Rabbit mAb detects a background band at 40 kDa in some cell lines. Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb may cross-react with phospho-IKKε. LC3B (E7X4S) XP® Rabbit mAb detects both type I and type II forms of LC3B. It weakly cross-reacts with LC3A by overexpression.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro143 of human LRSAM1, Gly15 of mouse Galectin-3/LGALS3, Val135 of human NDP52, residues near the carboxy terminus of human TAX1BP1 and human TBK1/NAK, residues near the amino terminus of human Atg16L1 and human LC3B, and synthetic phosphopeptides surrounding Ser278 of human Atg16L1 and Ser172 of human TBK1/NAK.
Background
Xenophagy provides an important defense against foreign pathogens, such as bacteria and viruses, by targeting them for degradation through autophagy (1-3). Several well-known pathogens can be targeted for degradation through xenophagy, including Salmonella typhimurium, Streptococcus pyogenes, and Mycobacterium tuberculosis, and loss of core autophagy genes can result in enhanced vulnerability to these infections. Pathogens are generally targeted to LC3B or other Atg8 family members localized on autophagosome membranes. Sequestosome 1/p62-like receptors (SLRs) function as cargo receptors through ubiquitin-mediated interactions along with LIR-domain interactions with Atg8 proteins on the autophagosome. The SLR TAX1BP1 is recruited to ubiquitylated Salmonella and plays a key role in xenophagy (4). Another SLR NDP52 has also been reported to be required for xenophagy of several pathogens (5,6). LRSAM1 is an E3 ubiquitin ligase with a leucine-rich repeat recognized by microbial ligands and recruits NDP52 (7). In addition, galectins are a family of cytosolic lectins that are associated with phagophores and bacteria. Galectin-3 is recruited to bacterial membranes and is important for protection against Mycobacterium tuberculosis (8). TBK1 is a serine/threonine kinase that plays a major role in controlling autophagy by targeting substrates in the SLR family and the maturation of autophagosomes. TBK1 is phosphorylated at Ser172 within its activation loop, which is required for its kinase activity (9). Atg16L1 is a component of the Atg12-Atg5 conjugation system required for autophagosome maturation (10,11). ULK1-mediated phosphorylation of Atg16L1 at Ser278 promotes xenophagy (12).
- Reggio, A. et al. (2020) Exp Cell Res 396, 112276.
- Levine, B. (2005) Cell 120, 159-62.
- Deretic, V. et al. (2013) Nat Rev Immunol 13, 722-37.
- Tumbarello, D.A. et al. (2015) PLoS Pathog 11, e1005174.
- Verlhac, P. et al. (2015) Cell Host Microbe 17, 515-25.
- Fan, S. et al. (2020) Int J Mol Sci 21, 2008. doi: 10.3390/ijms21062008.
- Huett, A. et al. (2012) Cell Host Microbe 12, 778-90.
- Kumar, S. et al. (2017) Autophagy 13, 1086-1087.
- Kishore, N. et al. (2002) J Biol Chem 277, 13840-7.
- Fujita, N. et al. (2008) Mol Biol Cell 19, 2092-100.
- Mizushima, N. et al. (2003) J Cell Sci 116, 1679-88.
- Alsaadi, R.M. et al. (2019) EMBO Rep 20, e46885.
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