Anti-ATG8 | Autophagy-related protein

• Background:

  ATG8 (Autophagy-related protein 8)is involved in degradation and recycling of intracellular components in a process of autophagy. ATG8 is a molecular autophagy marker inChlamydomonas reinhardtii(Pérez-Pérezet al. 2010, Plant Physiol. 152: 1874-88).

• Host:

  Rabbit

• Reactivity:

  Arabidopsis thaliana, Aponogeton madagascariensis, Chlamydononas reinhardtii,Chlorococcum dorsiventrale,Haematococcus lacustris, Nicotiana benthamiana, Populus trichocarpa, Solanum lycopersicum, Zea mays

• Not reactive in:

  Cuscuta chinensis,Symbiodiniumsp.

• Immunogen:

  Fragment of recombinant ATG8 fromChlamydomonas reinhardtii, UniProt:A8JB85

• Clonality:

  Polyclonal

• Applications:

  Immunolocalization (IL), Western blot (WB)

• Dilution:

  1 : 1000 (IL), 1 : 1000-1 : 2000 (WB)

• Purity:

  Serum

• Format:

  Lyophilized

• Reconstitution:

  For reconstitution add 50 µl of sterile water

• Molecular Weight:

  15.2 | 15 kDa

• Precautions:

  ForArabidopsis thalianathe signal obtained using ATG8 antibodies is cleaner in case of roots compare to leaf material. For best results please follow extraction protocol described inÁlvarezet al. (2012).ATG8 signal corresponds to the two bands of 17 kDa.Preparation of a cell extract fromArabidopsis thaliana:A. Plants were first subjected to autophagy activating conditions: nutrient (nitrogen or carbon) limitation or oxidative stress in order to activate this degradative process.B. Total protein extracts can be obtained as described byÁlvarez.Leaves are grinded in liquid nitrogen with a minimal volume of extraction buffer (100 mM Tris-HCl pH 8, 400 mM sucrose, 1 mM EDTA, 0.1 mM phenylmethylsulfonyl fluoride (PMSF), 10 mg/mL sodium deoxycholate, 10 µg/mL of leupeptin, 10 µg/mL of pepstatin A, 4% (v/v) protease inhibitor cocktail from Roche).C. Cell debris is removed by centrifuging at 500 g for 10 min at 4°C.Important note:It is recommendable to use bigger gels in order to get a better resolution of ATG8 bands. Midi-protean gels are better than mini-gels. There are 9 ATG8 isoforms and this antibody will likely recognizes all of them.For immunolocalization protocol, pleaseinquire.

• References & Citations:

  Liet al. (2024). Stress granules sequester autophagy proteins to facilitate plant recovery from heat stress. Nat Commun . 2024 Dec 30;15(1):10910. doi: 10.1038/s41467-024-55292-w.Liet al. (2024). Gamma-aminobutyric acid as a regulator of astaxanthin production in Haematococcus lacustris under salinity: Exploring physiology, signaling, autophagy, and multi-omics landscape. Bioresour Technol. 2024 Dec:413:131466.doi: 10.1016/j.biortech.2024.131466. Epub 2024 Sep 12.Michalaket al. (2024). Conserved autophagy and diverse cell wall composition: Unifying features of vascular tissues in evolutionarily distinct plants.Ann Bot. 2024 Apr 23;133(4):559-572. doi: 10.1093/aob/mcae015.Lanet al. (2024).Clathrin Light Chains negatively regulate plant immunity by hijacking the autophagy pathway. Plant Commun. 2024 Apr 30:100937.doi: 10.1016/j.xplc.2024.100937.Testiet al. (2024). The Phytophthora parasitica effector AVH195 interacts with ATG8, attenuates host autophagy, and promotes biotrophic infection. BMC Biol. 2024 Apr 29;22(1):100.Michalaket al. (2024). Conserved autophagy and diverse cell wall composition: Unifying features of vascular tissues in evolutionarily distinct plants. Ann Bot. 2024 Feb 7:mcae015. (immunofluorescence) doi: 10.1093/aob/mcae015.Llamaset al. (2023). In planta expression of human polyQ-expanded huntingtin fragment reveals mechanisms to prevent disease-related protein aggregation. Nat Aging. 2023 Nov;3(11):1345-1357.doi: 10.1038/s43587-023-00502-1.Miklaszewskaet al. (2023).CALEOSIN 1 interaction with AUTOPHAGY-RELATED PROTEIN 8 facilitates lipid droplet microautophagy in seedlings.Plant Physiol. 2023 Nov 22;193(4):2361-2380.doi: 10.1093/plphys/kiad471.Leeet al. (2023).Three consecutive cytosolic glycolysis enzymes modulate autophagic flux. Plant Physiol. 2023 Oct 26;193(3):1797-1815. doi: 10.1093/plphys/kiad439.Zhaoet al (2023) A non-canonical role of ATG8 in Golgi recovery from heat stress in plants. Nat Plants. 2023 May;9(5):749-765. doi: 10.1038/s41477-023-01398-w.Zenget al. (2023) The plant unique ESCRT component FREE1 regulates autophagosome closure. Nat Commun. 2023 Mar 30;14(1):1768. doi: 10.1038/s41467-023-37185-6.Sunet al. (2022) Genome of Paspalum vaginatum and the role of trehalose mediated autophagy in increasing maize biomass. Nat Commun. 2022;13(1):7731. Published 2022 Dec 13. doi:10.1038/s41467-022-35507-8Caoet al. (2022) Autophagic pathway contributes to low-nitrogen tolerance by optimizing nitrogen uptake and utilization in tomato. Hortic Res. 2022 Mar 23;9:uhac068. doi: 10.1093/hr/uhac068. PMID: 35669705; PMCID: PMC9164271.Sampernaet al (2022). Cyclopaldic Acid, the Main Phytotoxic Metabolite of Diplodia cupressi, Induces Programmed Cell Death and Autophagy in Arabidopsis thaliana. Toxins (Basel). 2022 Jul 11;14(7):474. doi: 10.3390/toxins14070474. PMID: 35878212; PMCID: PMC9325063.Zharovaet al. (2022) Role of Autophagy in Haematococcus lacustris Cell Growth under Salinity. Plants. 2022; 11(2):197. https://doi.org/10.3390/plants11020197 (immunofluorescence)Mishraet al. (2021) Interplay between abiotic (drought) and biotic (virus) stresses in tomato plants. Mol Plant Pathol. 2021 Dec 30. doi: 10.1111/mpp.13172. Epub ahead of print. PMID: 34970822.De Brasi-Velascoet al. (2021). Autophagy Is Involved in the Viability of Overexpressing Thioredoxin o1 Tobacco BY-2 Cells under Oxidative Conditions. Antioxidants. 2021; 10(12):1884.Sunet al. (2021) Mechanistic insights into an atypical interaction between ATG8 and SH3P2 in Arabidopsis thaliana. Autophagy. 2021 Oct 17:1-17. doi: 10.1080/15548627.2021.1976965. Epub ahead of print. PMID: 34657568.Liet al. (2021) The unfolded protein response plays dual roles in rice stripe virus infection through fine-tuning the movement protein accumulation. PLoS Pathog. 2021 Mar 4;17(3):e1009370. doi: 10.1371/journal.ppat.1009370. Epub ahead of print. PMID: 33662041.Liet al. (2021) Chloroplast proteotoxic stress-induced autophagy is involved in the degradation of chloroplast proteins in Chlamydomonas reinhardtii. Plant Cell Physiol. 2021 Feb 17:pcab029. doi: 10.1093/pcp/pcab029. Epub ahead of print. PMID: 33594417.Kazibweet al. (2020). TOR mediates the autophagy response to altered nucleotide homeostasis in a ribonuclease mutant. J Exp Bot. 2020 Sep 9;eraa410.doi: 10.1093/jxb/eraa410.Upadhyayaand Jagadeeshwar Rao (2019). Reciprocal regulation of photosynthesis and mitochondrial respiration by TOR kinase in Chlamydomonas reinhardtii. Plant Direct Volume 3, Issue 11.Shullet al. (2019). Anatase TiO2 nanoparticles induce autophagy and chloroplast degradation in thale cress (Arabidopsis thaliana). Environ Sci Technol. 2019 Jul 29. doi: 10.1021/acs.est.9b01648.Wojciechowskaet al. (2018). Autophagy counteracts instantaneous cell death during seasonal senescence of the fine roots and leaves in Populus trichocarpa. BMC Plant Biol. 2018 Oct 29;18(1):260. doi: 10.1186/s12870-018-1439-6. (immunolocalization)Chenet al. (2016). The role of nitric oxide signalling in response to salt stress in Chlamydomonas reinhardtii. Planta. 2016 Sep;244(3):651-69. doi: 10.1007/s00425-016-2528-0. Epub 2016 Apr 26.Gorovitset al. (2016). Tomato yellow leaf curl virus confronts host degradation by sheltering in small/midsized protein aggregates. Virus Res. 2016 Feb 2;213:304-13. doi: 10.1016/j.virusres.2015.11.020. Epub 2015 Dec 1.

• Storage Conditions:

  Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.

• CAS Number:

  9007-83-4