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Monoclonal Antibodies

Anti-ATG9 [Atg9 14F2 8B1]

Invented by Dr Sharon Tooze from The Francis Crick Institute
Invented at Cancer Research UK London Research Institute: Lincoln's Inn Fields
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Indirect immunouorscence on PFA fixed HEK293 cells labelled with hamster anti-Atg9 (Left) and rabbit anti-Atg9.
Western blot using HEK293 lysates either non-transfected (nt) or transfected (trans) with HA-tagged human Atg9. Arrowhead indicates position of human Atg9 at 105 KDa.
siRNA knockdown of human Atg9 (si) in Hela and Hek293 cells compared to siControl siRNA (siC) reveals specicity of hamster anti-Atg9 antibody.
IP Conditions: HEK293A cells. 1x15 cm dish. Cells lysed in 800 μl TNTE buffer (20 mM Tris pH 7.4, 150 mM NaCl, 5 mM EDTA and 0.4% Triton X-100) + protein inhibitors cocktail. Pi done using 10 μg of anti-ATG9A 14f2 8B1 antibody coupled to 50 μl of Protein A Dynabeads. o/n incubation of lysate with beads at 4°C. Elution with 50 μl SB 1x and 25 μl loaded for SDS-PAGE. Western blot: Blocking: 1x PBST with 5% non-fat dry milk Primary Ab dilution: Rabbit Anti ATG9A (STO215) 1/1000 in 1x PBST with 5% non-fat dry milk Secondary Ab dilution: HRP-Anti rabbit 1/5000 in 1x PBST with 5% non-fat dry milk
siRNA knockdown of HEK293A cells revealed with antiATG9A hamster 14f2 8B1 Orsi et al., MBoC 2012 23:1860-1873
Immunofluorescence in HEK293A cells revealed with anti-ATG9A hamster 14f2 8B1 Left: FM- full growth medium Right: ES- starved in EBSS Judith et al., J. Cell Biol. 2019 218: 1634–1652
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Catalogue Number 151525
Applications ELISA IHC IF IP WB
Antigen/Gene or Protein Targets Autophagy-related protein 9 (ATG9)
Reactivity Human, Mouse and Rat
Relevance Autophagy is an important process associated with infectious disease, neurodegeneration, cancer and the maintenance of cellular homeostasis. ATG9 is an essential component of the autophagy machinery and is found on autophagosomes. It is a multi-spanning membrane protein which makes it useful for sub-cellular localization studies. The detection of this protein provides important information about cell health and survival.
Host Hamster
Immunogen Conserved C-terminal synthetic peptide of Human ATG9 sequence
Positive Control Endogenous protein, tagged over expressed protein and sRNA-depleted HEK 293 cells,
Subclass IgG
Molecular Weight (kDa) 105
Myeloma Used P3X63Ag8.653
Recommended Growing Conditions Hybridoma Serum Free Medium + 7.5% FBS + 10 µg/ml gentamycin
Strain Armenian
Research Area Cancer, Cardiovascular, Cell Signaling & Signal Transduction, Metabolism, Neurobiology, Protein Degradation

References

There are 4 reference entries for this reagent.

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References: 4 entries

ATG9A shapes the forming autophagosome through Arfaptin 2 and phosphatidylinositol 4-kinase IIIβ

IHC IP

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Chan et al. 2009. Mol Cell Biol. 29(1):157-71. PMID: 18936157.

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Kinase-inactivated ULK proteins inhibit autophagy via their conserved C-terminal domains using an Atg13-independent mechanism.

Europe PMC ID: 18936157

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Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes.

IHC IP

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References: 4 entries

ATG9A shapes the forming autophagosome through Arfaptin 2 and phosphatidylinositol 4-kinase IIIβ

IHC IP

View  

Chan et al. 2009. Mol Cell Biol. 29(1):157-71. PMID: 18936157.

View  

Kinase-inactivated ULK proteins inhibit autophagy via their conserved C-terminal domains using an Atg13-independent mechanism.

View  

Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes.

IHC IP

View  

Add a reference

Inventor Information