All lanes : Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/2000 dilution (unpurified)

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : ACACA (Acetyl Coenzyme A Carboxylase) knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : A431 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 265 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab45174 observed at 265 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab45174 was shown to specifically react with Acetyl Coenzyme A Carboxylase in wild-type HAP1 cells as signal was lost in ACACA (Acetyl Coenzyme A Carboxylase) knockout cells. Wild-type and ACACA (Acetyl Coenzyme A Carboxylase) knockout samples were subjected to SDS-PAGE. Ab45174 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/2000 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Acetyl Coenzyme A Carboxylase with purified ab45174 at 1/400 dilution (2.06 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

All lanes : Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/1000 dilution (Purified)

Lane 1 : C2C12 (Mouse myoblasts myoblast) whole cell lysates
Lane 2 : Rat adrenal gland 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: 265 kDa
Observed band size: 265 kDa

Blocking/Diluting Buffer and concentration: 5% NFDM/TBST

We are unsure how to define the extra bands.

Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/1000 dilution (Purified) + A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 15 µg

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

Predicted band size: 265 kDa
Observed band size: 265 kDa

Blocking/Diluting Buffer and concentration: 5% NFDM/TBST

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomcah tissue sections labeling Acetyl Coenzyme A Carboxylase with purified ab45174 at 1/400 dilution (2.06 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling Acetyl Coenzyme A Carboxylase with purified ab45174 at 1/400 dilution (2.06 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

ab45174 (unpurified), at a dilution of 1/50, staining human Acetyl Coenzyme A Carboxylase in human liver by immunohistochemistry using paraffin embedded tissue. Heat mediated antigen retrieval was perfromed with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Acetyl Coenzyme A Carboxylase with ab45174 (unpurified) at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.