重组Anti-ATP citrate lyase抗体[EP704Y] (ab40793)
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
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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. -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P HumanIP HumanWB MouseRatHuman -
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 asab207504) -
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
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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
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.
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP704Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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Flow Cyt |
Human
|
ICC/IF |
Human
|
IHC-P |
Human
|
IP |
Human
|
WB |
Mouse
Rat
Human
|
All applications |
Common marmoset
|
应用 | Ab评论 | 说明 |
---|---|---|
IHC-P |
Use at an assay dependent concentration.
|
|
ICC/IF |
1/50.
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|
WB | (2) |
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.
|
说明 |
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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
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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
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数据库链接
- Entrez Gene: 47 Human
- Entrez Gene: 104112 Mouse
- Entrez Gene: 24159 Rat
- Omim: 108728 Human
- SwissProt: P53396 Human
- SwissProt: Q91V92 Mouse
- SwissProt: P16638 Rat
- Unigene: 387567 Human
see all -
别名
- ACL antibody
- Acly antibody
- ACLY_HUMAN antibody
see all
图片
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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.
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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) 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.
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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).
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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 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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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. -
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 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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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.
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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.
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.
数据表及文件
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SDS download
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Datasheet download
文献 (27)
ab40793 被引用在 27 文献中.
- Kagawa Y et al. FABP7 Regulates Acetyl-CoA Metabolism Through the Interaction with ACLY in the Nucleus of Astrocytes. Mol Neurobiol 57:4891-4910 (2020). PubMed: 32812201
- 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