Recombinant Anti-AMPK alpha 2 antibody [EP20772] (ab214425)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP20772] to AMPK alpha 2
- Suitable for: WB, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-AMPK alpha 2 antibody [EP20772]
See all AMPK alpha 2 primary antibodies -
Description
Rabbit monoclonal [EP20772] to AMPK alpha 2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293, HepG2 and HeLa whole cell lysates; Human skeletal muscle, fetal kidney and fetal liver tissue lysates. IP: HEK-293 whole cell lysate.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP20772 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab214425 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
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IP |
1/40.
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Notes |
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WB
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa). |
IP
1/40. |
Target
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Function
Catalytic 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. Regulates lipid synthesis by phosphorylating and inactivating lipid metabolic enzymes such as ACACA, ACACB, GYS1, HMGCR and LIPE; regulates fatty acid and cholesterol synthesis by phosphorylating acetyl-CoA carboxylase (ACACA and ACACB) and hormone-sensitive lipase (LIPE) enzymes, respectively. Regulates insulin-signaling and glycolysis by phosphorylating IRS1, PFKFB2 and PFKFB3. Involved in insulin receptor/INSR internalization (PubMed:25687571). AMPK stimulates glucose uptake in muscle by increasing the translocation of the glucose transporter SLC2A4/GLUT4 to the plasma membrane, possibly by mediating phosphorylation of TBC1D4/AS160. Regulates transcription and chromatin structure by phosphorylating transcription regulators involved in energy metabolism such as CRTC2/TORC2, FOXO3, histone H2B, HDAC5, MEF2C, MLXIPL/ChREBP, EP300, HNF4A, p53/TP53, SREBF1, SREBF2 and PPARGC1A. Acts as a key regulator of glucose homeostasis in liver by phosphorylating CRTC2/TORC2, leading to CRTC2/TORC2 sequestration in the cytoplasm. In response to stress, phosphorylates 'Ser-36' of histone H2B (H2BS36ph), leading to promote transcription. Acts as a key regulator of cell growth and proliferation by phosphorylating TSC2, RPTOR and ATG1/ULK1: in response to nutrient limitation, negatively regulates the mTORC1 complex by phosphorylating RPTOR component of the mTORC1 complex and by phosphorylating and activating TSC2. In response to nutrient limitation, promotes autophagy by phosphorylating and activating ATG1/ULK1. AMPK also acts as a regulator of circadian rhythm by mediating phosphorylation of CRY1, leading to destabilize it. May regulate the Wnt signaling pathway by phosphorylating CTNNB1, leading to stabilize it. Also phosphorylates CFTR, EEF2K, KLC1, NOS3 and SLC12A1. Plays an important role in the differential regulation of pro-autophagy (composed of PIK3C3, BECN1, PIK3R4 and UVRAG or ATG14) and non-autophagy (composed of PIK3C3, BECN1 and PIK3R4) complexes, in response to glucose starvation. Can inhibit the non-autophagy complex by phosphorylating PIK3C3 and can activate the pro-autophagy complex by phosphorylating BECN1. -
Sequence similarities
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
Contains 1 protein kinase domain. -
Domain
The AIS (autoinhibitory sequence) region shows some sequence similarity with the ubiquitin-associated domains and represses kinase activity. -
Post-translational
modificationsUbiquitinated.
Phosphorylated at Thr-172 by STK11/LKB1 in complex with STE20-related adapter-alpha (STRADA) pseudo kinase and CAB39. Also phosphorylated at Thr-172 by CAMKK2; triggered by a rise in intracellular calcium ions, without detectable changes in the AMP/ATP ratio. CAMKK1 can also phosphorylate Thr-172, but at much lower level. Dephosphorylated by protein phosphatase 2A and 2C (PP2A and PP2C). Phosphorylated by ULK1; leading to negatively regulate AMPK activity and suggesting the existence of a regulatory feedback loop between ULK1 and AMPK. Dephosphorylated by PPM1A and PPM1B at Thr-172 (mediated by STK11/LKB1). -
Cellular localization
Cytoplasm. Nucleus. In response to stress, recruited by p53/TP53 to specific promoters. - Information by UniProt
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Database links
- Entrez Gene: 5563 Human
- Omim: 600497 Human
- SwissProt: P54646 Human
- Unigene: 437039 Human
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Alternative names
- 5'-AMP-activated protein kinase catalytic subunit alpha-2 antibody
- AAPK2_HUMAN antibody
- ACACA kinase antibody
see all
Images
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All lanes : Anti-AMPK alpha 2 antibody [EP20772] (ab214425) at 1/5000 dilution
Lane 1 : His-tagged human AMPK alpha 1 recombinant protein (aa1-559)
Lane 2 : His-tagged human AMPK alpha 2 recombinant protein (aa1-552)
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 62 kDa
Observed band size: 62 kDa
Exposure time: 1 secondBlocking/Dilution: 5% NFDM/TBST.
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All lanes : Anti-AMPK alpha 2 antibody [EP20772] (ab214425) at 1/1000 dilution
Lane 1 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 62 kDa
Observed band size: 62 kDaBlocking/Dilution: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 30 seconds; Lane 3: 3 minutes.
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All lanes : Anti-AMPK alpha 2 antibody [EP20772] (ab214425) at 1/1000 dilution
Lane 1 : Human skeletal muscle tissue lysate
Lane 2 : Human fetal kidney tissue lysate
Lane 3 : Human fetal liver tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 62 kDa
Observed band size: 62 kDaBlocking/Dilution: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 5 seconds; Lane 3: 3 minutes.
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AMPK alpha 2 was immunoprecipitated from 0.35 mg of HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate with ab214425 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab214425 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1: HEK-293 whole cell lysate 10 µg (Input).
Lane 2: ab214425 IP in HEK-293 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214425 in HEK-293 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab214425 has been referenced in 1 publication.
- Mohseni R et al. Therapeutic effects of Chlorella vulgaris on carbon tetrachloride induced liver fibrosis by targeting Hippo signaling pathway and AMPK/FOXO1 axis. Mol Biol Rep 48:117-126 (2021). PubMed: 33296068