Anti-ATG9A antibody [SC67-05]
cat.: ET1610-71
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, ICC, IHC-P, IP
Clonality: Monoclonal
Clone number: SC67-05
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1 mg/mL.
Purification: Protein A affinity purified.
Molecular weight: 94 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human ATG9A aa 799-839 / 839.
Positive control: HepG2 cell lysates, Wild-type NCI-H1299 whole cell lysates, Hela, 293, rat brain tissue, mouse brain tissue, mouse heart tissue.
Subcellular location: Cytoplasmic vesicle, Golgi apparatus, Late endosome membrane, Endoplasmic reticulum membrane.
Recommended Dilutions:
  WB
  ICC
  IHC-P
  IP

1:500-1:2,000
1:50-1:200
1:50-1:200
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q7Z3C6 Human | Q68FE2 Mouse | Q5FWU3 Rat
Alternative names: APG9 autophagy 9-like 1 APG9 like 1 APG9-like 1 APG9L1 ATG9 ATG9 autophagy related 9 homolog A ATG9 autophagy related 9 homolog A (S. cerevisiae) ATG9A ATG9A_HUMAN Autophagy 9-like 1 protein Autophagy related protein 9A Autophagy-related protein 9A mATG9 MGD3208 OTTHUMP00000206046 OTTHUMP00000206048 OTTHUMP00000206049 OTTHUMP00000206062
Images
ET1610-71_1.jpg Fig1: Western blot analysis of ATG9A on HepG2 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-71, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
ET1610-71_2.jpg Fig2: All lanes: Western blot analysis of ATG9A with anti-ATG9A antibody [SC67-05] (ET1610-71) at 1:1,000 dilution.
Lane 1: Wild-type NCI-H1299 whole cell lysate (20 µg).
Lane 2: ATG9A knockout NCI-H1299 whole cell lysate (20 µg).

ET1610-71 was shown to specifically react with ATG9A in wild-type NCI-H1299 cells. No band was observed when ATG9A knockout sample was tested. Wild-type and ATG9A knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1610-71, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1610-71_3.jpg Fig3: ICC staining of ATG9A in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-71, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-71_4.jpg Fig4: ICC staining of ATG9A in 293 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-71, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-71_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-ATG9A antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-71, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-71_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-ATG9A antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-71, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-71_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-ATG9A antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-71, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.