Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

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.

• WB

Lab

Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

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

All lanes:

Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/1000 dilution

All lanes:

A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 266 kDa

Observed band size: 265 kDa

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• WB

Lab

Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

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.

All lanes:

Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/2000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

ACACA (Acetyl Coenzyme A Carboxylase) knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

A431 whole cell lysate at 20 µg

Predicted band size: 266 kDa

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• WB

Lab

Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

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

We are unsure how to define the extra bands.

All lanes:

Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (ab45174) at 1/1000 dilution

Lane 1:

C2C12 (Mouse myoblasts myoblast) whole cell lysates at 15 µg

Lane 2:

Rat adrenal gland lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 266 kDa

Observed band size: 265 kDa

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• WB

CiteAb

Western blot - Anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] (AB45174)

Acetyl Coenzyme A Carboxylase western blot using anti-Acetyl Coenzyme A Carboxylase antibody [EP687Y] ab45174. Publication image and figure legend from Karlas, A., Bèrre, S., et al., 2016, Nat Commun, PubMed 27177310.

ab45174 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab45174 please see the product overview.

Fatty acid synthesis requirement for CHIKV life cycle.(a) Impact of FASN or ACLY knockdown on CHIKV replication. Closed and open symbols indicate replicates from the primary screen and during validation, respectively. (b) Western blot showing silencing efficiency of siRNAs used in c. (c) Impact of FASN-, ACC- and ACLY-specific siRNAs on CHIKV replication (n=10 for each data set). (d) Confocal section of CHIKV replicon-infected HeLa cells labelled for FASN, dsRNA and 4,6-diamidino-2-phenylindole (DAPI; blue). Scale bar, 10 μm. (e) Co-localization analysis of cells labelled as in d and in Supplementary Fig. 3b, plotted as Pearson's coefficient per cell. Each symbol corresponds to a cell stack from three independent experiments (n=29 cells for FASN, 30 cells for ACC and 31 cells for ACLY); median values shown in red. (f) Effect of FASN (cerulenin, n=12 for each data set), ACC (TOFA, n=11 for each data set) and ACLY (BMS-303141 n=11 for each data set) inhibitors on CHIKV replication. (g) Real-time cell toxicity assay performed on HeLa cells (n=3 for each point). Excepted for b and d where representative images are shown and for g where the mean±s.d. is shown for each point of a representative experiment, all data represent the means±s.e.m. of three independent experiments analysed using one-way analysis of variance with Tukey's post test (*P<0.05; **P<0.01; ***P<0.001; NSP≥0.05). NS, not significant.

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