Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) cells labelling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and ACACA knockout HAP1 stained with ab269273 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab269273) (1x 10⁶ in 100μl at 0.008 μg/ml (1/74750 dilution)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HAP1 WT cells (black line) and HAP1-ACACA KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in wild-type HAP1 fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human breast carcinoma (PMID : 21415164). The section was incubated with ab269273 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

This image was produced using ab269273, the same clone but in a different formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% Triton X-100 permeabilized Hap1 WT and Hap1-ACACA KO cells labelling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 μg/ml dilution (Green). Image showing cytoplasmic staining in Hap1 WT cell line. ab7291 Anti-alpha Tubulin antibody [DM1A] was used to counterstain tubulin at 1/1000 dilution (Magenta). The nuclear counterstain was DAPI (Blue).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T cells labelling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in 293T cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 μg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue labeling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human endometrial carcinoma. The section was incubated with ab269273 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (Mouse myoblasts myoblast) cells labelling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 cells labelling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) antibody at 1/1000 2 μg/ml dilution (Green). Confocal image showing cytoplasmic staining in C2C12 cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^®594) was used to counterstain tubulin at 1/200 2.5 μg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) at 1/1000 2 μg/ml dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse lung (PMID : 17521700). The section was incubated with ab269273 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling Acetyl Coenzyme A carboxylase alpha with ab269273 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat lung (PMID : 17521700). The section was incubated with ab269273 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

• IP

Lab

Immunoprecipitation - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Acetyl Coenzyme A carboxylase alpha was immunoprecipitated from 0.35 mg C2C12 (mouse myoblasts myoblast), whole cell lysate 10 μg with ab269273 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269273 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : C2C12 whole cell lysate 10 μg.

Lane 2 : ab269273 IP in C2C12 whole cell lysate 10 μg.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab269273 in C2C12 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

All lanes:

Immunoprecipitation - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] (<a href='/en-us/products/primary-antibodies/acetyl-coenzyme-a-carboxylase-alpha-antibody-epr23235-147-ab269273'>ab269273</a>)

Predicted band size: 265 kDa

false

• IP

Lab

Immunoprecipitation - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] - BSA and Azide free (AB272704)

Acetyl Coenzyme A carboxylase alpha was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell), whole cell lysate 10 μg with ab269273 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269273 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 293T whole cell lysate 10 μg.

Lane 2 : ab269273 IP in 293T whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab269273 in 293T whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269273).

All lanes:

Immunoprecipitation - Anti-Acetyl Coenzyme A carboxylase alpha antibody [EPR23235-147] (<a href='/en-us/products/primary-antibodies/acetyl-coenzyme-a-carboxylase-alpha-antibody-epr23235-147-ab269273'>ab269273</a>)

Predicted band size: 265 kDa

false