Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-ATP citrate lyase antibody [EP704Y] - BSA and Azide free (ab227996)

Overview

  • Product name
    Anti-ATP citrate lyase antibody [EP704Y] - BSA and Azide free
    See all ATP citrate lyase primary antibodies
  • Description
    Rabbit monoclonal [EP704Y] to ATP citrate lyase - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, ICC/IF, Flow Cyt, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Common marmoset
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human ATP citrate lyase aa 1050 to the C-terminus (C terminal). The exact sequence is proprietary.
    (Peptide available as ab207504)

  • Positive control
    • WB: HeLa cell lysate. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: Jurkat cell lysate.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab227996 is a PBS only buffer version of ab40793, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab40793 for information on protocols, dilutions, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Properties

Applications

Our Abpromise guarantee covers the use of ab227996 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. PubMed: 19727777
WB Use at an assay dependent concentration. Detects a band of approximately 125 kDa (predicted molecular weight: 122 kDa).Can be blocked with ATP citrate lyase peptide (ab207504).

Target

  • Function
    ATP-citrate synthase is the primary enzyme responsible for the synthesis of cytosolic acetyl-CoA in many tissues. Has a central role in de novo lipid synthesis. In nervous tissue it may be involved in the biosynthesis of acetylcholine.
  • Sequence similarities
    In the N-terminal section; belongs to the succinate/malate CoA ligase beta subunit family.
    In the C-terminal section; belongs to the succinate/malate CoA ligase alpha subunit family.
    Contains 1 ATP-grasp domain.
  • Post-translational
    modifications
    ISGylated.
    Acetylated at Lys-540, Lys-546 and Lys-554 by KAT2B/PCAF. Acetylation is promoted by glucose and stabilizes the protein, probably by preventing ubiquitination at the same sites. Acetylation promotes de novo lipid synthesis. Deacetylated by SIRT2.
    Ubiquitinated at Lys-540, Lys-546 and Lys-554 by UBR4, leading to its degradation. Ubiquitination is probably inhibited by acetylation at same site.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • ACL antibody
    • Acly antibody
    • ACLY_HUMAN antibody
    • ATP citrate (pro-S) lyase antibody
    • ATP citrate lyase antibody
    • ATP citrate synthase antibody
    • ATP-citrate (pro-S-)-lyase antibody
    • ATP-citrate synthase antibody
    • ATPcitrate synthase antibody
    • ATPCL antibody
    • Citrate cleavage enzyme antibody
    • CLATP antibody
    • OTTHUMP00000164773 antibody
    see all

Images

  • This WB data was generated using the same tni-ATP citrate lyase antibody clone, EP704Y, in a different buffer formulation (cat# ab40793).

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: ATP citrate lyase knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab40793 observed at 125 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab40793 was shown to specifically react with ATP citrate lyase in wild-type HAP1 cells as signal was lost in ATP citrate lyase knockout cells. Wild-type and ATP citrate lyase knockout samples were subjected to SDS-PAGE. ab40793 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.

  • ab40793 (purified) at 1:20 dilution (1.5μg) immunoprecipitating ATP citrate lyase in HeLa whole cell lysate.

    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate,10μg
    Lane 2 (+): ab40793 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40793 in HeLa whole cell lysate. 

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40793).

  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ATP citrate lyase with purified ab40793 at 1:30 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor ® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40793).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human clear cell carcinoma of kidney tissue sections labeling ATP citrate lyase with Purified ab40793 at 1:100 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40793).

  • Unpurified ab40793 at 1/40 immunoprecipitating ATP citrate lyase in HeLa (human cervix adenocarcinoma) whole cell lysate.

    Lane 1 (input): HeLa (human cervix adenocarcinoma) whole cell lysate 10μg

    Lane 2 (+): ab40793 + HeLa (human cervix adenocarcinoma) whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40793 in HeLa (human cervix adenocarcinoma) whole cell lysate

    For western blotting, ab40793 at 1/1000 dilution and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40793).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ATP citrate lyase with purified ab40793 at 1/50. Cells were fixed with 100% methanol. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
    Control: PBS only.
    Nuclear counter stain: DAPI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40793).

  • Overlay histogram showing HeLa cells stained with unpurified 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 (ab40793, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40793).

  • This ICC/IF data was generated using the same tni-ATP citrate lyase antibody clone, EP704Y, in a different buffer formulation (cat# ab40793).

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ATP citrate lyase with ab40793 at 1/500. Cells were fixed with 100% methanol. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
    Control: PBS only.
    Nuclear counter stain: DAPI.

References

This product has been referenced in:
  • Chu DT  et al. C57BL/6J mice as a polygenic developmental model of diet-induced obesity. Physiol Rep 5: (2017). WB ; Mouse . Read more (PubMed: 28400497) »
  • Zhang C  et al. Cullin3-KLHL25 ubiquitin ligase targets ACLY for degradation to inhibit lipid synthesis and tumor progression. Genes Dev 30:1956-70 (2016). WB ; Human . Read more (PubMed: 27664236) »
See all 14 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab227996.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up