使用敲除细胞株进行验证重组RabMAb

重组Anti-ATP citrate lyase抗体[EP704Y] (ab40793)

Reviews (2)Q&A (3)References (26)

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
  • Suitable for: IHC-P, ICC/IF, WB, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

概述

  • 产品名称

    Anti-ATP citrate lyase抗体[EP704Y]
    参阅全部 ATP citrate lyase 一抗

  • 描述

    兔单克隆抗体[EP704Y] to ATP citrate lyase

  • 宿主

    Rabbit

  • 特异性

    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.

  • 经测试应用

    适用于: IHC-P, ICC/IF, WB, Flow Cyt, IPmore details

  • 种属反应性

    与反应: Mouse, Rat, Human
    预测可用于: Common marmoset

  • 免疫原

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

  • 阳性对照

    • 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

  • 常规说明

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

应用

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.

应用 Ab评论 说明
IHC-P Use at an assay dependent concentration.
ICC/IF 1/50.
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.

靶标

  • 功能

    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.

  • 序列相似性

    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.

  • 翻译后修饰

    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.

  • 细胞定位

    Cytoplasm.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • 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.

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

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

文献 (26)

发表研究结果有使用 ab40793？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab40793 被引用在 26 文献中.

  • 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). 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). PubMed: 30452972
  • Xue D  et al. Quantitative proteomic analysis of sperm in unexplained recurrent pregnancy loss. Reprod Biol Endocrinol 17:52 (2019). PubMed: 31288842
  • Favara DM  et al. ADGRL4/ELTD1 Silencing in Endothelial Cells Induces ACLY and SLC25A1 and Alters the Cellular Metabolic Profile. Metabolites 9:N/A (2019). PubMed: 31775252
  • Chen Y  et al. ATP-citrate lyase is an epigenetic regulator to promote obesity-related kidney injury. FASEB J 33:9602-9615 (2019). PubMed: 31150280
View all Publications for this product

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1-2 of 2 Abreviews

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

提交于 Jun 05 2014

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

提交于 Oct 02 2012

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