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Anti-ACLY Antibody EP704Y
Also for ACLY (NM_001096)
|A synthetic peptide corresponding to residues in the C-term of human ATP citrate lyase was used as immunogen.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IF, IP, FC
||ICC/IF: Use a concentration of 5 ug/ml; IHC-P: 1:100; WB: 1:1000 - 1:5000; FC: 1:100; IP: Use at an assay dependent concentration
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|IgG fraction (Protein A or G Sepharose)
|Homo sapiens ATP citrate lyase (ACLY), transcript variant 1|
|ACL; ATPCL; CLATP|
|ATP citrate lyase (ACL) is the primary enzyme responsible for the synthesis of cytosolic acetyl-CoA in many tissues. The enzyme is a tetramer of four identical subunits. It catalyzes the formation of acetyl-CoA and oxaloacetate from citrate and CoA with a concomitant hydrolysis of ATP to ADP and phosphate. One of these products, acetyl-CoA, serves several important biosynthetic pathways, including lipogenesis and cholesterogenesis. In nervous tissue, ATP citrate-lyase may be involved in the biosynthesis of acetylcholine. NDPK has been found to phosphorylate ACL and insulin to increase phosphorylation of ACL (2).|
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Western blot - ATP citrate lyase antibody [EP704Y]; Anti-ATP citrate lyase antibody [EP704Y] at 1/5000 dilution + Hela cell lysate.Predicted band size : 122 kDa.Observed band size : 122 kDa.
Immunocytochemistry/ Immunofluorescence - Anti-ATP citrate lyase antibody [EP704Y]; ICC/IF image of ab40793 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was Alexa Fluor? 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunoprecipitation - Anti-ATP citrate lyase antibody [EP704Y]; AMPK gamma 1 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 10µg of Rabbit monoclonal [Y308] to AMPK gamma 1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). .The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with .Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) .Band: 122kDa: ATP citrate lyase; non specific - 60kDa: We are unsure as to the identity of this extra band.
Flow Cytometry - ATP citrate lyase antibody [EP704Y]; Overlay histogram showing HeLa cells stained with ab40793 (red line). The cells were fixed with 80% methanol (5 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 for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed.