Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-ATP citrate lyase antibody [EP704Y] (ab40793)

Reviews (2)Q&A (3)References (22)

Overview

  • Product name

    Anti-ATP citrate lyase antibody [EP704Y]
    See all ATP citrate lyase primary antibodies

  • Description

    Rabbit monoclonal [EP704Y] to ATP citrate lyase

  • Host species

    Rabbit

  • Specificity

    ab40793 recognises ATP citrate lyase (ACL). The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human, Common marmoset

  • Immunogen

    Synthetic peptide 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; NIH/3T3; rat lung; C6 lysates. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: Jurkat cell lysate; HeLa. IHC: Human clear cell carcinoma of kidney

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab40793 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
ICC/IF 1/50.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. PubMed: 19727777

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
WB 1/10000. Detects a band of approximately 125 kDa (predicted molecular weight: 122 kDa).Can be blocked with ATP citrate lyase peptide (ab207504).

For unpurified use at 1/1000 - 1/5000.
Flow Cyt 1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/100. 
IP 1/20.

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

    see all

  • 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

  • Western blot - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Western blot - Anti-ATP citrate lyase antibody [EP704Y] (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.

    Unpurified 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Immunocytochemistry/ Immunofluorescence - Anti-ATP citrate lyase antibody [EP704Y] (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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP citrate lyase antibody [EP704Y] (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.
  • Flow Cytometry - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Flow Cytometry - Anti-ATP citrate lyase antibody [EP704Y] (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).
  • Western blot - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Western blot - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    All lanes : Anti-ATP citrate lyase antibody [EP704Y] (ab40793) at 1/50000 dilution (purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 122 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Immunoprecipitation - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)

    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.

  • Western blot - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Western blot - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    All lanes : Anti-ATP citrate lyase antibody [EP704Y] (ab40793) at 1/10000 dilution (purified)

    Lane 1 : Rat lung lysates
    Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 122 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Western blot - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Anti-ATP citrate lyase antibody [EP704Y] (ab40793) at 1/5000 dilution (unpurified) + HeLa cell lysate

    Predicted band size: 122 kDa
    Observed band size: 122 kDa

  • Flow Cytometry - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Flow Cytometry - Anti-ATP citrate lyase antibody [EP704Y] (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.

  • Immunoprecipitation - Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
    Immunoprecipitation - Anti-ATP citrate lyase antibody [EP704Y] (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 ab131366 VeriBlot for IP (HRP) was used for detection at 1/10000.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

References

This product has been referenced in:

  • Jin F  et al. HIF-1a-induced miR-23a~27a~24 cluster promotes colorectal cancer progression via reprogramming metabolism. Cancer Lett 440-441:211-222 (2019). Read more (PubMed: 30393198) »
  • Shi Z  et al. Silibinin inhibits endometrial carcinoma via blocking pathways of STAT3 activation and SREBP1-mediated lipid accumulation. Life Sci 217:70-80 (2019). Read more (PubMed: 30452972) »
See all 22 Publications for this product

Publishing research using ab40793? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Western blot abreview for Anti-ATP citrate lyase antibody [EP704Y]

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
80 µg
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Sample
Marmoset (common) Tissue lysate - whole (brain)
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Jun 05 2014

Question

What's the concentration?

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Answer

Vielen Dank für Ihren Anruf.

Leider ermitteln wir für diesen Antikörper die Konzentration nicht, so dass ich Ihnen leider in diesem Fall nicht weiterhelfen kann.

Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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Western blot abreview for Anti-ATP citrate lyase antibody [EP704Y]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (Head)
Loading amount
40 µg
Specification
Head
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Oct 02 2012

Question

How are you?

I am almost ready to submit the blots showing it worked. Whatexactlyshould I send you so I could get two new antibodies for free?

Read More
Answer


Please log into your Abcam account, and than open the datasheet for the antibody you would like to write a review about. On top of the datasheet, you will find five tags : Datasheet, specific references, Abreview, Scientific Support, Protocols.

You have to click (you guessed it) on the Abreview section, and just follow the Submit an Abreviewlink.

You will find a drop- down menu which is pretty self explainingfrom which you have to choose your used application, species, blocking conditions, etc. An image is always welcome as well, so feel free to upload one.

However, to inform us that this is a 100% Test-discount Review, please add one of the discount codes into the "additional information "section at the end of the review. Submit it, and voila, you are ready to order anantibody for free!

Read More

Question

,


Thank you very much!

The first antibody is :

Anti-Histone H4 (acetyl K12) antibody (ab61238)
Which appears to also work on WB Drosophila.


The second is:

Anti-ATP citrate lyase antibody [EP704Y] (ab40793)
Which appears to work on WB in Drosophila - I want to run more test to make sure.



What does the discount means exactly? How can I use it?


Bw

Read More
Answer

Expiration date: 08. September 2012

ab40793 -DISCOUNT CODE: xxxx


ab61238- DISCOUNT CODE: xxxx

I am very pleased to hear you accepted our offer and tested ab40793 und ab61238 in drosophila. This code will give you: 1 freeprimary antibody per codebefore the expiration date. To redeem this offer, please submit an Abreview fordrosophila for each antibody and include the aprobriatecode in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:
1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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