Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172)抗体(ab23875)
Key features and details
- Rabbit polyclonal to AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172)
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
- Reacts with: Human
- Isotype: IgG
选择批间可重复性更高的重组抗体
- 研究可靠 —— 各批次间结果一致且可重复
- 长期批量供应 —— 采用重组技术,可实现快速生产
- 首次实验即可成功 —— 经过大量验证确认了特异性
- 符合伦理标准 —— 产品不含动物成分
概述
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产品名称
Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172)抗体 -
描述
兔多克隆抗体to AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) -
宿主
Rabbit -
特异性
ab23875 recognises the phosphorylated forms of AMPK alpha 1 (T183) and AMPK alpha 2 (T172).
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经测试应用
适用于: WB, IHC-P, Flow Cyt, ICC/IFmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide corresponding to Human AMPK alpha 1 (phospho T183). Also within AMPK alpha 2 (phospho T172).
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阳性对照
- Insulin treated CHO T cells, Insulin treated 3T3L1, Metformin treated L6 myoblast cells. Metformin treated HepG2 cells (10 mM for 24 hr).
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常规说明
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
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. -
存储溶液
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA -
Concentration information loading...
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纯度
Immunogen affinity purified -
纯化说明
ab23875 was purified from rabbit serum by sequential epitope specific chromatography. The antibody has been negatively preadsorbed using a non phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non phosphorylated AMPK. The final product is generated by affinity chromatography using a AMPK derived peptide that is phosphorylated at threonine 172. -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab23875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 |
1/1000. Predicted molecular weight: 62 kDa.
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IHC-P |
1/10 - 1/50.
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Flow Cyt |
1/20.
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ICC/IF |
1/250.
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说明 |
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WB
1/1000. Predicted molecular weight: 62 kDa. |
IHC-P
1/10 - 1/50. |
Flow Cyt
1/20. |
ICC/IF
1/250. |
靶标
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细胞定位
AMPK alpha 2: Cytoplasm. Nucleus. In response to stress, recruited by p53/TP53 to specific promoters. -
数据库链接
- Entrez Gene: 5562 Human
- Entrez Gene: 5563 Human
- Omim: 600497 Human
- Omim: 602739 Human
- SwissProt: P54646 Human
- SwissProt: Q13131 Human
- Unigene: 43322 Human
- Unigene: 437039 Human
图片
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All lanes : Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) antibody (ab23875)
Lane 1 : Lysates prepared from HepG2 cells left unstimulated with 3% BSA-TBST buffer and no peptide
Lane 2 : Lysates prepared from HepG2 cells stimulated with
Metformin with 3% BSA-TBST buffer and no peptide
Lane 3 : Lysates prepared from HepG2 cells stimulated with
Metformin with 3% BSA-TBST buffer and the non-phosphopeptide corresponding to the immunogen
Lane 4 : Lysates prepared from HepG2 cells stimulated with
Metformin with 3% BSA-TBST buffer and a
generic phospho-threonine-containing peptide
Lane 5 : Lysates prepared from HepG2 cells stimulated with
Metformin with 3% BSA-TBST buffer and the phosphopeptide
immunogen
Lane 6 : Lysates prepared from HepG2 cells stimulated with
Metformin and treated with lambda
phosphatase with 3% BSA-TBST buffer
Secondary
All lanes : goat F(ab’)2 anti rabbit
IgG HRP conjugate
Predicted band size: 62 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 2 hoursLysates were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF, treated or not with lambda phosphatase, blocked with a 3% BSA-TBST buffer for one hour at room temperature, incubated with relevant peptides (see below) and incubated with the AMPK alpha 1/2 [pT 172] antibody for two hours at room temperature in 3% BSA-TBST buffer.
After washing, membranes were incubated with goat F(ab’)2 antirabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal™ method.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue sections labeling AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) with ab23875 (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) antibody (ab23875) diluted in 3% BSA-PBS at a dilution of 1/20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
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Immunocytochemistry/ Immunofluorescence analysis of 70% confluent log phase MDA-MB-231 cells labeling AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) with ab23875. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) antibody (ab23875) at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) secondary antibody, Alexa Fluor® 488 conjugate at a dilution of 1/2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with mountant with DAPI. F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin, 1/300. Panel d is a merged image showing Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
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Flow Cytometry analysis of MDA-MB-231 cells labeling AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) with ab23875. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) antibody (ab23875, red) or with rabbit isotype control (pink) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody at a dilution of 1/400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (72)
ab23875 被引用在 72 文献中.
- Zhao X et al. Deciphering the Mechanism of Bushen Huoxue Decotion on Decidualization by Intervening Autophagy via AMPK/mTOR/ULK1: A Novel Discovery for URSA Treatment. Front Pharmacol 13:794938 (2022). PubMed: 35140613
- Yu X et al. Empagliflozin Inhibits Hepatic Gluconeogenesis and Increases Glycogen Synthesis by AMPK/CREB/GSK3β Signalling Pathway. Front Physiol 13:817542 (2022). PubMed: 35299662
- Cheng L et al. Rutin-activated adipose tissue thermogenesis is correlated with increased intestinal short-chain fatty acid levels. Phytother Res 36:2495-2510 (2022). PubMed: 35445769
- Wu J et al. Ferulic acid ameliorates acetaminophen-induced acute liver injury by promoting AMPK-mediated protective autophagy. IUBMB Life 74:880-895 (2022). PubMed: 35514074
- Jin C et al. Higenamine Attenuates Doxorubicin-Induced Cardiac Remodeling and Myocyte Apoptosis by Suppressing AMPK Activation. Front Cell Dev Biol 10:809996 (2022). PubMed: 35602605