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ATG13 phospho S318 Antibody

Rabbit Polyclonal
NCI Collaboration
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Western blot using Rockland's affinity purified anti-ATG13 pS318 antibody shows detection of phosphorylated ATG13 in 293T cells engineered to coexpress Ulk1 and Atg13 (Ulk1 + Atg13).  In the left lane was loaded kinase-dead hypophosphorylated Ulk1-K46A mutant + ATG13.  The right lane contains the 293T Ulk1 + ATG13 lysate and shows detection at approximately 57 kDa. The antibody was purified and resolved by SDS-PAGE, then transferred to nitrocellulose membrane. The membrane was blocked with 5% Blotto (p/n B501-0500) and probed with the primary antibody at 1µg/mL overnight at 4°C. After washing, the membrane was probed with Goat Anti-Rabbit HRP secondary 1:5000  in detection buffer (p/n MB-070) for 45 minutes at room temperature. In collaboration with Charles Dorsey at Eli Lilly, Indianapolis, IN and John Cleveland at Scripps, Jupiter, FL.
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25 µL
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ATG13 phospho S318 Antibody Properties

Anti-ATG13 pS318 (RABBIT) Antibody - 600-401-C49S
Target Species
Known Cross Reactivity
ELISA : 1:25,000-1:175,000
Western Blot : 1:1000
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping Condition
Dry Ice
1.1 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
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ATG13 phospho S318 Antibody Description

ATG13 is a target of the TOR kinase signaling pathway that regulates autophagy through the control of the phosphorylation status of ATG13 and ULK1 through their stable complex, and the regulation of ATG13-ULK1-RB1CC1. ATG13 also forms a stable complex with FIP200. Ulk1 phosphorylates ATG13 on S318 and promotes its release to damaged mitochondria. Autophagy is a normal process in eukaryotes required for turnover of cellular components during starvation and stress. It plays an essential role in cellular differentiation, cell death and aging. Defects in this evolutionarily conserved process may contribute to certain human diseases such as cancer, neurodegenerative diseases, muscular disorders and pathogen infections. ATG13 is one of several ATG genes required for autophagosome formation in mammalian cells. mTOR interacts with this complex in a nutrient dependent manner and phosphorylates Atg13 and ULK1.
Autophagy-related protein 13, KIAA0652
This affinity purified antibody was prepared by repeated immunizations with a synthetic peptide corresponding to the region near S318 of ATG13.
Immunogen Type
Post Translational Modification
Storage Condition
Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
Application Note
This affinity purified antibody has been tested for use in ELISA and by western blot.  Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 56.6 kDa in size corresponding to human phosphorylated ATG13 protein by western blotting in the appropriate stimulated tissue or cell lysate or extract. 
This affinity-purified antibody is directed against the phosphorylated form of human ATG13 protein at the pS318 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification.  Antiserum was first purified against the phosphorylated form of the immunizing peptide.  The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.  Reactivity occurs against human ATG13 pS318 protein and the antibody is specific for the phosphorylated form of the protein.   Reactivity with non-phosphorylated human ATG13 is minimal by ELISA and western blot.  A BLAST analysis was used to suggest cross reactivity with ATG13 from human based on 100% sequence homology with the immunogen.  Reactivity against homologues from other sources is not known.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Specific Reference
Egan, D. F., Chun, M. G., Vamos, M., Zou, H., Rong, J., Miller, C. J., ... & Shaw, R. J. (2015). Small Molecule Inhibition of the Autophagy Kinase ULK1 and Identification of ULK1 Substrates. Molecular cell, 59(2), 285-297. Joo JH, Dorsey FC, Joshi A, Hennessy-Walters KM, Rose KL, McCastlain K, Zhang J, Iyengar R, Jung CH, Suen DF, Steeves MA, Yang CY, Prater SM, Kim DH, Thompson CB, Youle RJ, Ney PA, Cleveland JL, Kundu M. (2011) Hsp90-Cdc37 Chaperone Complex Regulates Ulk1- and Atg13-Mediated Mitophagy. Mol Cell. Aug 19;43 (4):572-85. Nazio F, Strappazzon F, Antonioli M, Bielli P, Cianfanelli V, Bordi M, Gretzmeier C, Dengjel J, Piacentini M, Fimia GM, Cecconi F. (2013) mTOR inhibits autophagy by controlling ULK1 ubiquitylation, self-association and function through AMBRA1 and TRAF6. Nat Cell Biol. Apr;15 (4):406-16. Stanton MJ, Dutta S, Zhang H, Polavaram NS, Leontovich AA, Hönscheid P, Sinicrope FA, Tindall DJ, Muders MH, Datta K. (2013) Autophagy Control by the VEGF-C/NRP-2 Axis in Cancer and Its Implication for Treatment Resistance. Cancer Res. Jan 1;73 (1):160-71.
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