重组Anti-AMPK gamma 1抗体[Y307] (ab32382)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y307] to AMPK gamma 1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-AMPK gamma 1抗体[Y307]
参阅全部 AMPK gamma 1 一抗 -
描述
兔单克隆抗体[Y307] to AMPK gamma 1 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-Pmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human AMPK gamma 1 aa 1-100. The exact sequence is proprietary.
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表位
ab32382 reacts with an epitope located in the N terminal region of AMPK gamma 1. -
阳性对照
- Jurkat cell lysate and HeLa cell lysate, human skeletal muscles
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y307 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32382于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (1) |
1/1000 - 1/2000. Predicted molecular weight: 38 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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说明 |
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WB
1/1000 - 1/2000. Predicted molecular weight: 38 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
靶标
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功能
AMP/ATP-binding subunit of AMP-activated protein kinase (AMPK), an energy sensor protein kinase that plays a key role in regulating cellular energy metabolism. In response to reduction of intracellular ATP levels, AMPK activates energy-producing pathways and inhibits energy-consuming processes: inhibits protein, carbohydrate and lipid biosynthesis, as well as cell growth and proliferation. AMPK acts via direct phosphorylation of metabolic enzymes, and by longer-term effects via phosphorylation of transcription regulators. Also acts as a regulator of cellular polarity by remodeling the actin cytoskeleton; probably by indirectly activating myosin. Gamma non-catalytic subunit mediates binding to AMP, ADP and ATP, leading to activate or inhibit AMPK: AMP-binding results in allosteric activation of alpha catalytic subunit (PRKAA1 or PRKAA2) both by inducing phosphorylation and preventing dephosphorylation of catalytic subunits. ADP also stimulates phosphorylation, without stimulating already phosphorylated catalytic subunit. ATP promotes dephosphorylation of catalytic subunit, rendering the AMPK enzyme inactive. -
序列相似性
Belongs to the 5'-AMP-activated protein kinase gamma subunit family.
Contains 4 CBS domains. -
结构域
The AMPK pseudosubstrate motif resembles the sequence around sites phosphorylated on target proteins of AMPK, except the presence of a non-phosphorylatable residue in place of Ser. In the absence of AMP this pseudosubstrate sequence may bind to the active site groove on the alpha subunit (PRKAA1 or PRKAA2), preventing phosphorylation by the upstream activating kinase STK11/LKB1.
The CBS domains mediate binding to AMP, ADP and ATP. 2 sites bind either AMP or ATP, whereas a third site contains a tightly bound AMP that does not exchange. Under physiological conditions AMPK mainly exists in its inactive form in complex with ATP, which is much more abundant than AMP. -
翻译后修饰
Phosphorylated by ULK1 and ULK2; leading to negatively regulate AMPK activity and suggesting the existence of a regulatory feedback loop between ULK1, ULK2 and AMPK. - Information by UniProt
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数据库链接
- Entrez Gene: 5571 Human
- Omim: 602742 Human
- SwissProt: P54619 Human
- Unigene: 530862 Human
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别名
- 5' AMP activated protein kinase gamma 1 subunit antibody
- 5' AMP activated protein kinase subunit gamma 1 antibody
- 5''-AMP-activated protein kinase subunit gamma-1 antibody
see all
图片
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All lanes : Anti-AMPK gamma 1 antibody [Y307] (ab32382) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : PRKAG1 (AMPK gamma 1) knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 38 kDaLanes 1 - 3: Merged signal (red and green). Green - ab32382 observed at 38 kDa. Red - loading control, Mouse anti Vinculin, observed at 125 kDa.
ab32382 was shown to recognize AMPK gamma 1 in wild-type HAP1 cells as signal was lost at the expected MW in PRKAG1 (AMPK gamma 1) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PRKAG1 (AMPK gamma 1) knockout samples were subjected to SDS-PAGE. Ab32382 and Mouse anti-Vinculin (ab9484 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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All lanes : Anti-AMPK gamma 1 antibody [Y307] (ab32382) at 1/2000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Predicted band size: 38 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
This image shows paraffin embedded human skeletal muscle stained with 1/250 ab23282.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)This image is courtesy of an anonymous AbreviewAll lanes : Anti-AMPK gamma 1 antibody [Y307] (ab32382) at 1/2000 dilution
Lane 1 : HeLa whole cell lystate - Control Si
Lane 2 : HeLa whole cell lystate - treated with AMPK gamma 1 siRNA A for 36 hours
Lane 3 : HeLa whole cell lystate - treated with AMPK gamma 1 siRNA B for 36 hours
Secondary
All lanes : HRP-conjugated Donkey polyclonal to rabbit IgG at 1/3000 dilution
Predicted band size: 38 kDa
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 (4)
ab32382 被引用在 4 文献中.
- Puustinen P et al. DNA-dependent protein kinase regulates lysosomal AMP-dependent protein kinase activation and autophagy. Autophagy 16:1871-1888 (2020). PubMed: 31983282
- Ren XM et al. Atorvastatin Alleviates Experimental Diabetic Cardiomyopathy by Regulating the GSK-3ß-PP2Ac-NF-?B Signaling Axis. PLoS One 11:e0166740 (2016). PubMed: 27851811
- Head SA et al. Antifungal drug itraconazole targets VDAC1 to modulate the AMPK/mTOR signaling axis in endothelial cells. Proc Natl Acad Sci U S A 112:E7276-85 (2015). PubMed: 26655341
- Preuss KD et al. Inactivation of protein-phosphatase 2A causing hyperphosphorylation of autoantigenic paraprotein targets in MGUS/MM is due to an exchange of its regulatory subunits. Int J Cancer 135:2046-53 (2014). PubMed: 24676687