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Synthetic peptide within Human Acetyl Coenzyme A carboxylase alpha aa 1-100 (N terminal). The exact sequence is proprietary.
Mouse and rat: We have preliminary internal testing data to indicate this antibody may not react with these species, Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab109368 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000. Predicted molecular weight: 266 kDa.
For unpurified use at 1/1000- 1/10000.
|IHC-P||1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Flow Cyt||1/100 - 1/500.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/100 - 1/250.|
Lanes 1 - 4: Merged signal (red and green). Green - ab109368 observed at 265 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab109368 was shown to specifically react with Acetyl Coenzyme A carboxylase alpha 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. Ab109368 and ab130007 (Mouse anti-Vinculin 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.
Immunocytochemistry/ Immunofluorescence analysis of 293 (Human embryonic kidney epithelial cell) cells labeling Acetyl Coenzyme A carboxylase alpha with Purified ab109368 at 1:250 dilution (2.1μg/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). 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.
Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Sonic Hedgehog with purified ab109368 at 1:100 dilution (5 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - 90% methanol. Unlabeled control - Rabbit monoclonal IgG (Black). Cells without incubation with primary antibody and secondary antibody (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling Acetyl Coenzyme A carboxylase alpha with purified ab109368 at 1:500 dilution (1.05 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using citrate Buffer, pH6.0. Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.
Overlay histogram showing SH-SY5Y cells stained with unpurified ab109368 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109368, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunofluorescent staining of 293 cells using unpurified ab109368 at 1/100 dilution
Immunohistochemical analysis of paraffin-embedded skeletal muscle tissue using unpurified ab109368 at 1/250 dilution.
Immunohistochemical analysis of paraffin-embedded brain tissue using unpurified ab109368 at 1/250 dilution.
ab109368 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"