Atg12

Manufactured by Santa Cruz Biotechnology

Sourced in United States

ATG12 is a protein involved in the autophagy process. It plays a critical role in the formation of autophagosomes, which are double-membrane vesicles that engulf cellular components for degradation and recycling. ATG12 conjugates with ATG5 to form a complex that is essential for the initiation and expansion of the autophagosome membrane.

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Lab products found in correlation

Hoechst, by Thermo Fisher Scientific (2 mentions) Etoposide, by Merck Group (2 mentions) Doxorubicin, by Merck Group (2 mentions) Chloroquine, by Merck Group (2 mentions) N acetylcysteine, by Merck Group (2 mentions) Phospho histone h3, by Cell Signaling Technology (2 mentions) Quinacrine, by Merck Group (2 mentions) Cleaved parp, by Abcam (2 mentions) Phospho p44 42 mapk p erk, by Thermo Fisher Scientific (2 mentions) P44 42 mapk erk1 2, by Thermo Fisher Scientific (2 mentions) Abc232, by Merck Group (2 mentions) 6 aminonicotinamide, by Merck Group (2 mentions) 1 13c glucose, by Cambridge Isotopes (2 mentions) 1 14c glucose and 6 14c glucose, by PerkinElmer (2 mentions) β actin, by Novus Biologicals (1 mentions) Becn1, by Santa Cruz Biotechnology (1 mentions) Cdkn1a, by Cell Signaling Technology (1 mentions) Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions) Caspase 3, by Cell Signaling Technology (1 mentions) Phosphorylated akt ser473, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Anti-ATG12 (8 citations)

6 protocols using atg12

Immunoblotting and Immunofluorescence Analysis of Autophagy Markers

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Commercial antibodies include: phospho-Histone H3 (Cell Signaling Technology (CST) 9701), cleaved caspase-3 Alexa Fluor 488 (CST 9669S), phospho-p44/42 MAPK (P-ERK) (Invitrogen 44–680G), p44/42 MAPK (ERK1/2) (Invitrogen 13–6200), ATG12 (CST 2010), ATG7 (Santa Cruz Biotechnology sc8668), p62 (Progen Biotechnik GP62C), tubulin (Sigma T6199), p53 (DO1, Calbiochem OP43), cleaved PARP (CST 9451), G6PD (Abcam ab993) LC3 5F10 for IF (Axxora NT0231-00). For immunobloting, we utilized an LC3 antibody which has been described previously and is now commercially available (EMD Millipore ABC232) (35 (link)). Chemicals include: chloroquine (CQ), quinacrine (Q), 6-aminonicotinamide (6-AN), N-acetyl Cysteine (NAC), etoposide (Et) and doxorubicin (DX), all from Sigma-Aldrich, staurosporine (STS, EMD Chemicals), 1-¹³C glucose (Cambridge Isotopes), 1-¹⁴C glucose and 6-¹⁴C glucose (Perkin Elmer), and Hoechst (Invitrogen).

Salas E., Roy S., Marsh T., Rubin B, & Debnath J. (2015). Oxidative Pentose Phosphate Pathway Inhibition Is A Key Determinant of Antimalarial Induced Cancer Cell Death. Oncogene, 35(22), 2913-2922.

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Antibody Detection for Autophagy Signaling

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Antibodies were obtained from: GeneTex, Inc (TGM2, #GTX111702), Cell Signaling (CDKN1A, #2947; LC3B, #2775; ATG12, #D88H11; ATG5, #D5F5U; BECN1, #D40C5), Santa Cruz Biotechnology (TP53, DO-1, #sc-261), Millipore (β-actin, #MAB1501), and Novus Biologicals (SQSTM1/p62, 2C11, #H00008878-M01).

Yeo S.Y., Itahana Y., Guo A.K., Han R., Iwamoto K., Nguyen H.T., Bao Y., Kleiber K., Wu Y.J., Bay B.H., Voorhoeve M, & Itahana K. (2016). Transglutaminase 2 contributes to a TP53-induced autophagy program to prevent oncogenic transformation. eLife, 5, e07101.

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Proteomic Analysis of Cellular Stress Response

Cited 1 time

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Lysates of 5 × 10⁶ cells were prepared using Mammalian Cell Extraction Kits (Abcam, Cambridge, MA). HSP60, RPTOR, Atg4 (#ab108322, Abcam, Cambridge, MA), Atg12 (sc-68884, Santa Cruz Biotechnology), LC3 (#PA1-16930, ThermoFischer Scientific), phosphorylated ERK (#9106), phosphorylated Ser473-Akt (#9721), phosphorylated RPTOR (#2083), Bax (#2774), cytochrome c (#12963), caspase3 (#9662), ubiquitin (#3936), and actin (#4967, Cell Signaling Technology, Danvers, MA) antibodies were used for probing the designated proteins in lysates using immunoblotting protocols. In some experiments, protein aggregates in 100 μg cell lysates were isolated after ultrahigh centrifugation at 10,000 g at 4 ℃ for 30 min, as previously described^{20 (link)}. For positive controls, 100 μg cell lysates were mixed with equal volume of ice-cold acetone. Pellets were separated by electrophoresis and stained using Coomassie blue, and RPTOR levels in the pellet was probed using immunoblotting protocols. In addition, HSP60 and RPTOR immunoprecipitates were isolated for immunoblotting probed using ubiquitin antibody.

Lian W.S., Ko J.Y., Chen Y.S., Ke H.C., Wu S.L., Kuo C.W, & Wang F.S. (2018). Chaperonin 60 sustains osteoblast autophagy and counteracts glucocorticoid aggravation of osteoporosis by chaperoning RPTOR. Cell Death & Disease, 9(10), 938.

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HeLa Cell Autophagy Induction by S1

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Human cervical cancer cells (HeLa cells) were obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA) and cultured in Iscove’s modified Dulbecco’s media (IMDM) (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% (v/v) FBS (Thermo Fisher Scientific) at 37°C with 5% CO₂. When S1 was used to treat the HeLa cells, the concentration was 10 μM/L in IMDM. The antibody against Bcl-2 (1:500, rabbit) and LC3 (1:500, rabbit) was purchased from Abcam (Cambridge, MA, USA). The antibody against Bax (1:100, mouse) and Atg 12 (1:100, rabbit) was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). The antibody against caspase-3 (1:500, rabbit) was purchased from Cell Signaling (Beverly, MA, USA). The antibody against Beclin (1:100, mouse) was purchased from BD Biosciences (San Jose, CA, USA).

Zhou L., Kong Q., Zhang Y., Yang W., Yan S., Li M., Wang Y., Zhou Y., Yu H, & Han L. (2018). Glucose deprivation promotes apoptotic response to S1 by enhancing autophagy in human cervical cancer cells. Cancer Management and Research, 10, 6195-6204.

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Autophagy Induction in Cancer Cells

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ATO, a kind gift from Intas Pharmaceuticals Ltd, Ahmedabad, India, was used in the study. Daunorubicin, Cytarabine, Bafilomycin A1, Hydroxychloroquine, Bay11-7086 was procured from Sigma, St. Louis, USA. Antibodies used included those against Actin, ATG12, Beclin1, p62, p65 (Santa Cruz, CA, USA), LC3 (Cell Signaling Technology Inc, Massachusetts, USA), TLR2 (BD Pharmingen, New Jersey, USA) anti-mouse and anti-rabbit secondary antibodies conjugated with horseradish peroxidase (Cell Signaling Technology Inc., Massachusetts, USA) and with alexaflour 488 and 594 (Invitrogen, California, USA) were used for western blotting and immunofluorescence.

Ganesan S., Palani H.K., Lakshmanan V., Balasundaram N., Alex A.A., David S., Venkatraman A., Korula A., George B., Balasubramanian P., Palakodeti D., Vyas N, & Mathews V. (2019). Stromal cells downregulate miR-23a-5p to activate protective autophagy in acute myeloid leukemia. Cell Death & Disease, 10(10), 736.

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Immunoblotting and Immunofluorescence Analysis of Autophagy Markers

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Salas E., Roy S., Marsh T., Rubin B, & Debnath J. (2015). Oxidative Pentose Phosphate Pathway Inhibition Is A Key Determinant of Antimalarial Induced Cancer Cell Death. Oncogene, 35(22), 2913-2922.

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