PRODUCT CODE: ET1610-71

ATG9A Recombinant Rabbit Monoclonal Antibody [SC67-05] (ET1610-71)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

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.
  • 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.
  • Western blot analysis of ATG9A on Wild-type NCI-H1299 whole cell lysate and ATG9A knockout NCI-H1299 whole cell lysate. Proteins at 20 µg per lane were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-71, 1/1,000) was used in 5% NFDM/TBST 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. Rabbit anti-GAPDH (ET1601-4) loading control were incubated for 1 hour at room temperature at 1/10,000 dilution respectively.<br />
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The cell line was provided by Ubigene Biosciences Co., Ltd., Guangzhou, China.
  • 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).
  • 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).
  • 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.
  • 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.
  • 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.
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.

Applications

  • WB

  • ICC

  • IHC-P

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

ATG9A Recombinant Rabbit Monoclonal Antibody [SC67-05] (ET1610-71)

Immunogen

Synthetic peptide within c-terminal human atg9a.

Host

Rabbit

Positive Control

HepG2 cell lysates, Hela, 293, rat brain tissue, mouse brain tissue, mouse heart tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SC67-05

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

94 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

ATG9A

SYNONYMS

APG9 autophagy 9-like 1 antibody; APG9 like 1 antibody; APG9-like 1 antibody; APG9L1 antibody; ATG9 antibody; ATG9 autophagy related 9 homolog A antibody; ATG9 autophagy related 9 homolog A (S. cerevisiae) antibody; ATG9A antibody; ATG9A_HUMAN antibody; Autophagy 9-like 1 protein antibody; Autophagy related protein 9A antibody; Autophagy-related protein 9A antibody; mATG9 antibody; MGD3208 antibody; OTTHUMP00000206046 antibody; OTTHUMP00000206048 antibody; OTTHUMP00000206049 antibody; OTTHUMP00000206062 antibody

SEQUENCE SIMILARITIES

Belongs to the ATG9 family.

SUBCELLULAR LOCATION

Cytoplasmic vesicle, Golgi apparatus, Late endosome membrane, Endoplasmic reticulum membrane.

FUNCTION

Autophagy, a process that results in the lysosomal-dependent degradation of cytosolic compartments, is carried out by the autophagosome, which is a double-membrane vesicle whose formation is catalyzed by several autophagy-related gene (Atg) proteins. Atg9a (autophagy-related protein 9A), also known as APG9-like 1, is a 839 amino acid, multi-pass membrane protein that localizes to the pre-autophagosomal structure (PAS). Isolation membranes are suggested to originate from the PAS, enwrapping cytoplasmic components to form a double membrane autophagosome, which then fuses with the vacuole. Ubiquitously expressed in human adult tissues, Atg9a cycles between the Golgi and endosomes and, with the autophagosome-specific marker LC3, plays a critical role in starvation-induced autophagosome formation. Three isoforms of Atg9a exist as a result of alternative splicing events.