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| Product Name | ATP citrate lyase Recombinant Rabbit Monoclonal Antibody |
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| Antibody Type | Primary Antibodies |
| Product description | ATP-citrate synthase, also designated ATP-citrate lyase or citrate cleavage enzyme, is a cytoplasmic homotetramer belonging to the succinate/malate CoA ligase family. The gene coding for this protein maps against chromosome 17q12-q21. ATP-citrate synthase catalyses the formation of acetyl-CoA and oxaloacetate from citrate and CoA. This product, Acetyl-CoA, is necessary for both fatty acid and cholesterol biosynthesis. ATP citrate-lyase is important in the biosynthesis of acetylcholine in nervous tissue. |
| Immunogen | Synthetic peptide within C-terminal human ATP citrate lyase. |
| Clonality | monoclonal |
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| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:1000-1:2000 ICC/IF:1:1000 IHC:1:200-1:1000 FC:1:50-1:200 IP:1-2μg/sample | |
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | ACLY |
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| Gene Synonyms | ACL ATPCL CLATP |
| Gene Full Name | ATP citrate lyase |
| Molecular Weight(MW) | 121kDa |
| Cellular Localization | Cytoplasm, cytosol. |
WB
Western blot analysis of ATP citrate lyase on different lysates with Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate, Lane 2: NIH/3T3 cell lysate, Lane 3: C6 cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 10 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling ATP citrate lyase with Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling ATP citrate lyase with Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
FC
Flow cytometric analysis of HeLa cells labeling ATP citrate lyase. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
ATP citrate lyase was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with Rabbit anti-ATP citrate lyase antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-ATP citrate lyase antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: NIH/3T3 cell lysate (input), Lane 2:Rabbit anti-ATP citrate lyase antibody IP in NIH/3T3 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-ATP citrate lyase antibody in NIH/3T3 cell lysate, Blocking/Dilution buffer: primary antibody dilution,Exposure time: 2 seconds.| Positive Control | A549, MCF-7, Hela, CRC, mouse kidney tissue, human thyroid tissue, human breast carcinoma tissue, mouse thyroid tissue. |
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| Application Notes | WB:1:1000-1:2000 ICC/IF:1:1000 IHC:1:200-1:1000 FC:1:50-1:200 IP:1-2μg/sample |
| Form | Liquid |
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| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
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