Western blot analysis of ACC1 was performed using membrane enriched extracts (30 ug lysate) of A549 (Lane 1), K-562 (Lane 2), HeLa (Lane 3),PC-3 (Lane 4), HEK-293 (Lane 5), Caco-2 (Lane 6) and Hep G2 (Lane 7). The blots were probed with Anti-ACC1 Mouse monoclonal Antibody (Product # 43-7100, 1-3 µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/ml, 1:2500 dilution). A 265 kDa band corresponding to ACC1 were observed across the cell lines tested. Along with the desired bands, non-specific bands at 120 kDa were observed across cell lines, except PC-3. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and HiMark™ Pre-stained Protein Standard (Product # LC5699). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant protein derived from the C-terminus of human Acetyl CoA Carboxylase 1 (ACC-1) protein (accession # Q13085, NP_942131), which is identical to chimpanzee. This protein is also 99% similar to Rhesus monkey, 96% similar to pig and bovine and 95% similar to mouse and rat.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Acetyl-CoA carboxylase (ACC) is a complex multifunctional enzyme system. ACC is a biotin-containing enzyme which catalyzes the carboxylation of acetyl-CoA to malonyl-CoA, the rate-limiting step in fatty acid synthesis. There are two ACC forms, alpha and beta, encoded by two different genes. ACC-alpha is highly enriched in lipogenic tissues. The enzyme is under long term control at the transcriptional and translational levels and under short term regulation by the phosphorylation/dephosphorylation of targeted serine residues and by allosteric transformation by citrate or palmitoyl-CoA. Multiple alternatively spliced transcript variants divergent in the 5' sequence and encoding distinct isoforms have been found for this gene.
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