Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Monoamine Oxidase A/MAO-A with Purified ab126751 at 1:100 dilution (1.5 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling Monoamine Oxidase A/MAO-A with Purified ab126751 at 1:400 dilution (0.38 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Monoamine Oxidase A/MAO-A with Purified ab126751 at 1:400 dilution (0.38 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue sections labeling Monoamine Oxidase A/MAO-A with Purified ab126751 at 1:400 dilution (0.38 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

All lanes : Anti-Monoamine Oxidase A/MAO-A antibody [EPR7101] (ab126751) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : MAOA (Monoamine Oxidase A) knockout HAP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 60 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab126751 observed at 60 kDa. Red - loading control, ab8245, observed at 0 kDa.

ab126751 was shown to recognize Monoamine Oxidase A in wild-type HAP1 cells as signal was lost at the expected MW in MAOA (Monoamine Oxidase A) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and MAOA (Monoamine Oxidase A) knockout samples were subjected to SDS-PAGE. Ab126751 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Anti-Monoamine Oxidase A/MAO-A antibody [EPR7101] (ab126751) at 1/10000 dilution (Purified) + Mouse brain lysates at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 60 kDa
Observed band size: 60 kDa

All lanes : Anti-Monoamine Oxidase A/MAO-A antibody [EPR7101] (ab126751) at 1/1000 dilution (Purified)

Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 2 : Rat brain lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 60 kDa
Observed band size: 60 kDa

Anti-Monoamine Oxidase A/MAO-A antibody [EPR7101] (ab126751) at 1/2000 dilution (unpurified) + Mouse brain cortex tissue lysate - whole at 20 µg

Secondary
HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/2000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 60 kDa
Observed band size: 60 kDa


Exposure time: 1 minute

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ab126751 (unpurified), at 1/50 dilution, staining Monoamine Oxidase A/MAO-A in paraffin embedded Human colon tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab126751 (purified) staining Monoamine Oxidase A/MAO-A in the human cell line HepG2 (human hepatocellular carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/20. 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)

All lanes : Anti-Monoamine Oxidase A/MAO-A antibody [EPR7101] (ab126751) at 1/1000 dilution (unpurified)

Lane 1 : HepG2 lysate
Lane 2 : NCI-H460 lysate
Lane 3 : Fetal liver lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 60 kDa

ab126751 (unpurified), at 1/50 dilution, staining Monoamine Oxidase A/MAO-A in paraffin embedded Human kidney tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Equilibrium disassociation constant (K_D)
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