Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP704Y] to ATP citrate lyase - BSA and Azide free
  • Suitable for: IHC-P, IP, ICC/IF, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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 & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab207504)

  • Positive control

    • WB: HeLa cell lysate. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: Jurkat cell lysate.

  • General notes

    Ab227996 is the carrier-free version of ab40793. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab227996 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

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

    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.

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

    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 - Anti-ATP citrate lyase antibody [EP704Y] - BSA and Azide free (ab227996)
    Flow Cytometry - Anti-ATP citrate lyase antibody [EP704Y] - BSA and Azide free (ab227996)
    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) - Anti-ATP citrate lyase antibody [EP704Y] - BSA and Azide free (ab227996)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase antibody [EP704Y] - BSA and Azide free (ab227996)
    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).

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

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

    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).

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

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-ATP citrate lyase antibody [EP704Y] - BSA and Azide free (ab227996)
    Immunocytochemistry/ Immunofluorescence - Anti-ATP citrate lyase antibody [EP704Y] - BSA and Azide free (ab227996)

    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.

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

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