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RabMAb

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

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: ICC/IF, IHC-P, WB, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Common marmoset
  • 免疫原

    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)

  • 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

    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.

    This product is a recombinant rabbit monoclonal antibody.

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
ICC/IF 1/50.
IHC-P 1/100. 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).

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
  • 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

  • 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.

  • 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.

  • 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 secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • 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).
  • 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.
  • 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 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.

  • 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

  • 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.

  • 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 as the secondary antibody at 1/10000.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

文献

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:N/A (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

客户评价及客户问答

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
Username

Abcam user community

Verified customer

提交于 Jun 05 2014

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...

Read More
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
Username

Abcam user community

Verified customer

提交于 Oct 02 2012


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. Read More

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 f...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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