Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (AB223528)

Unpurified ab108338, at 1/100, staining ATG9A in paraffin-embedded Human colon tissue by Immunohistochemistry.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (AB223528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human thyroid carcinoma tissue sections labeling ATG9A with Purified ab108338 at 1 : 50 dilution (4.12 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (AB223528)

Unpurified ab108338 staining ATG9A in the human cell line HepG2 (human hepatocellular carcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/40. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (AB223528)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATG9A with Purified ab108338 at 1 : 100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor^® 488) was used as the secondary antibody at 1 : 1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (AB223528)

Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATG9A with purified ab108338 at 1 : 20 dilution (10 µg/ml) (red). Cells were fixed with 100% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1 : 2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (AB223528)

Unpurified ab108338 at 1/50 dilution, staining ATG9A in HepG2 cells by Immunofluorescence.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

• IP

Lab

Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (AB223528)

ab108338 (purified) at 1 : 20 dilution (2μg) immunoprecipitating ATG9A in HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate.

Lane 1 (input) : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab108338 & HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab108338 in HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate

For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1 : 1000 dilution. No band in input lane is due to the boiled lysates
Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

All lanes:

Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] (<a href='/en-us/products/primary-antibodies/atg9a-antibody-epr24502-ab108338'>ab108338</a>)

Predicted band size: 94 kDa

false

• WB

Lab

Western blot - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (AB223528)

This western blot data was generated using the same anti-ATG9A clone (EPR2450(2)] in a different buffer formulation (cat# ab108338).

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : ATG9A knockout HAP1 cell lysate (20 μg)
Lane 3 : HepG2 cell lysate (20 μg)
Lane 4 : A375 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab108338 observed at 100 and 130 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab108338 was shown to specifically react with ATG9A when ATG9A knockout samples were used. Wild-type and ATG9A knockout samples were subjected to SDS-PAGE. ab108338 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-ATG9A antibody [EPR2450(2)] (<a href='/en-us/products/primary-antibodies/atg9a-antibody-epr24502-ab108338'>ab108338</a>)

Predicted band size: 94 kDa

false