Revision 1
Cell Signaling Technology

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Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
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UniProt ID:

#O75143

Entrez-Gene Id:

9776

Product Includes Quantity Reactivity MW(kDa) Isotype
Phospho-Atg13 (Ser355) (E4D3T) Rabbit mAb 46329 100 µl H M R 72 Rabbit IgG
Atg13 (E1Y9V) Rabbit mAb 13468 100 µl H 72 Rabbit IgG

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation but has also been associated with a number of physiological processes including development, differentiation, neurodegeneration, infection, and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and referred to as autophagy-related (Atg) genes.

Atg13/Apg13 was originally identified in yeast as a constitutively expressed protein that was genetically linked to Atg1/Apg1, a protein kinase required for autophagy (4). Overexpression of Atg1 suppresses the defects in autophagy observed in Atg13 mutants (4). Autophagy requires a direct association between Atg1 and Atg13, and is inhibited by TOR-dependent phosphorylation of Atg13 under high-nutrient conditions (5). Similarly, mammalian Atg13 forms a complex with the Atg1 homologues ULK1/2, along with FIP200, which localizes to autophagic isolation membranes and regulates autophagosome biogenesis (6-8). mTOR phosphorylates both Atg13 and ULK1, suppressing ULK1 kinase activity and autophagy (7-9). ULK1 can directly phosphorylate Atg13 at a yet unidentified site, presumably to promote autophagy (7,8). Additional studies suggest that Atg13 and FIP200 can function independently of ULK1 and ULK2 to induce autophagy through an unknown mechanism (10).
ULK1-dependent phosphorylation of Atg13 at Ser355, which corresponds to Ser318 of isoform 2 of Atg13, leads to the recruitment of Atg13 to damaged mitochondria, enabling efficient mitophagy (11).

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
  2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
  3. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
  4. Funakoshi, T. et al. (1997) Gene 192, 207-13.
  5. Kamada, Y. et al. (2000) J Cell Biol 150, 1507-13.
  6. Ganley, I.G. et al. (2009) J Biol Chem 284, 12297-305.
  7. Hosokawa, N. et al. (2009) Mol Biol Cell 20, 1981-91.
  8. Jung, C.H. et al. (2009) Mol Biol Cell 20, 1992-2003.
  9. Kim, J. et al. (2011) Nat Cell Biol 13, 132-41.
  10. Alers, S. et al. (2011) Autophagy 7, 1423-33.
  11. Joo, J.H. et al. (2011) Mol Cell 43, 572-85.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PhosphoPlus is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #36105

    PhosphoPlus® Atg13 (Ser355) Antibody Duet

    PhosphoPlus® Atg13 (Ser355) Antibody Duet: Image 1 Expand Image
    Western blot analysis of extracts from various cell lines using Atg13 (E1Y9V) Rabbit mAb.
    PhosphoPlus® Atg13 (Ser355) Antibody Duet: Image 2 Expand Image
    Western blot analysis of extracts from A-673 cells, untreated (-) or treated as indicated with the ULK1 inhibitor SBI-0206965 #29089 (50 μM, 2hr; +), Earle's Balanced Salt Solution (EBSS, 2 hr; +), or Torin 1 #14379 (250 nM, 2 hr; +), using Phospho-Atg13 (Ser355) (E4D3T) Rabbit mAb (upper), Atg13 (E1Y9V) Rabbit mAb #13468 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    PhosphoPlus® Atg13 (Ser355) Antibody Duet: Image 3 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Atg13 (hAtg13-Myc/DDK; +), using Atg13 (E1Y9V) Rabbit mAb (upper) or Myc-Tag (71D10) Rabbit mAb #2278 (lower).
    PhosphoPlus® Atg13 (Ser355) Antibody Duet: Image 4 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Atg13 protein (hAtg13-Myc/DDK; +) or mouse ULK1 (mULK1; +), using Phospho-Atg13 (Ser355) (E4D3T) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    PhosphoPlus® Atg13 (Ser355) Antibody Duet: Image 5 Expand Image
    Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Atg13 siRNA I #12043 (+), using Atg13 (E1Y9V) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The Atg13 (E1Y9V) Rabbit mAb confirms silencing of Atg13 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
    PhosphoPlus® Atg13 (Ser355) Antibody Duet: Image 6 Expand Image
    Immunoprecipitation of phospho-Atg13 (Ser355) protein from A-673 cell extracts treated with Torin 1 #14379 (250 nM, 2 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Atg13 (Ser355) (E4D3T) Rabbit mAb. Western blot analysis was performed using Phospho-Atg13 (Ser355) (E4D3T) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
    PhosphoPlus® Atg13 (Ser355) Antibody Duet: Image 7 Expand Image
    Immunoprecipitation of Atg13 from MCF7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (lane 2) or Atg13 (E1Y9V) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Atg13 (E1Y9V) Rabbit mAb.
    PhosphoPlus® Atg13 (Ser355) Antibody Duet: Image 8 Expand Image
    Confocal immunofluorescent analysis of PANC-1 cells, untreated (left) or nutrient-starved with Earle's balanced salt solution (3 hr; right), using Atg13 (E1Y9V) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).