Phospho-AMPK alpha 1 (S496) Recombinant Rabbit Monoclonal Antibody [SD0810] - BSA and Azide free
Catalog# HA750300
Phospho-AMPK alpha 1 (S496) Recombinant Rabbit Monoclonal Antibody [SD0810] - BSA and Azide free
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WB
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IP
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IHC-Fr
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Human
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Mouse
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ET1612-72
含抗保成分
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unconjugated
Overview
Product Name
Phospho-AMPK alpha 1 (S496) Recombinant Rabbit Monoclonal Antibody [SD0810] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Ser496 of Human AMPK alpha 1 aa 471-520 / 559.
Species Reactivity
Human, Mouse
Validated Applications
WB, IP, IHC-Fr
Molecular Weight
Predicted band size: 64 kDa
Positive Control
SiHa cell lysate, HUVEC cell lysate, K-562 cell lysate, mouse hippocampus tissue, mouse cerebral cortex tissue.
Conjugation
unconjugated
Clone Number
SD0810
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:1,000
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IP
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Use at an assay dependent concentration.
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IHC-Fr
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1:100
Target
Function
AMPK (5'-AMP-activated protein kinase) is a heterotrimeric complex comprising a catalytic α subunit and regulatory β and γ subunits. It protects cells from stresses that cause ATP depletion by switching off ATP-consuming bio-synthetic pathways. AMPK is activated by high AMP and low ATP through a mechanism involving allosteric regulation, promotion of phosphorylation by an upstream protein kinase known as AMPK kinase, and inhibition of dephosphorylation. Activated AMPK can phosphorylate and regulate in vivo hydroxy-methylglutaryl-CoA reductase and acetyl-CoA carboxylase, which are key regulatory enzymes of sterol synthesis and fatty acid synthesis, respectively. AMPKα1 (5'-AMP-activated protein kinase catalytic subunit alpha-1), also known as PRKAA1, is a 559 amino acid protein that belongs to the CAMK Ser/Thr protein kinase family and protein kinase superfamily. Highly phosphorylated, AMPKα1 exists as two alternatively spliced isoforms and is encoded by a gene that maps to human chromosome 5p13.1.
Background References
1. Chang TJ et al. Glucagon-like peptide-1 prevents methylglyoxal-induced apoptosis of beta cells through improving mitochondrial function and suppressing prolonged AMPK activation. Sci Rep 6:23403 (2016).
2. Wang YG & Yang TL Liraglutide reduces fatty degeneration in hepatic cells via the AMPK/SREBP1 pathway. Exp Ther Med 10:1777-1783 (2015).
Sequence Similarity
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
Post-translational Modification
Ubiquitinated.; Phosphorylated at Thr-183 by STK11/LKB1 in complex with STE20-related adapter-alpha (STRADA) pseudo kinase and CAB39. Also phosphorylated at Thr-183 by CAMKK2; triggered by a rise in intracellular calcium ions, without detectable changes in the AMP/ATP ratio. CAMKK1 can also phosphorylate Thr-183, but at a much lower level. Dephosphorylated by protein phosphatase 2A and 2C (PP2A and PP2C). 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. Dephosphorylated by PPM1A and PPM1B.
Subcellular Location
Nucleus, Cytoplasm.
Synonyms
5 AMP activated protein kinase alpha 1catalytic subunit antibody
5 AMP activated protein kinase catalytic alpha 1 chain antibody
5' AMP activated protein kinase catalytic subunit alpha 1 antibody
5'-AMP-activated protein kinase catalytic subunit alpha-1 antibody
AAPK1 antibody
AAPK1_HUMAN antibody
ACACA kinase antibody
acetyl CoA carboxylase kinase antibody
AI194361 antibody
AI450832 antibody
ExpandImages
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Western blot analysis of Phospho-AMPK alpha 1 (S496) on different lysates with Rabbit anti-Phospho-AMPK alpha 1 (S496) antibody (HA750300) at 1/5,000 dilution.
Lane 1: SiHa cell lysate
Lane 2: HUVEC cell lysate
Lane 3: K-562 cell lysate
Lysates/proteins at 15 µg/Lane.
Predicted band size: 64 kDa
Observed band size: 64 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750300) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Western blot analysis of Phospho-AMPK alpha 1(S496) on HUVEC cell lysates.
Lane 1: HUVEC cells, whole cell lysate, 10ug/lane
Lane 2/3: HUVEC cells treated with UV(2 hours recovery), whole cell lysates, 10ug/lane
Lane 4: HUVEC cells treated with UV(2 hours recovery), then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane
All lanes :
Anti-Phospho-AMPK alpha 1(S496) antibody (HA750300) at 1:500 dilution. Anti-GAPDH antibody (ET1601-4) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution.
Predicted band size:64 kDa
Observed band size:64 kDa
Blocking and diluting buffer: 5% BSA.
Exposure time: 2 minutes 14 seconds -
Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Phospho-AMPK alpha 1 (S496) with Rabbit anti-Phospho-AMPK alpha 1 (S496) antibody (HA750300).
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (HA750300, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner. -
Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Phospho-AMPK alpha 1 (S496) with Rabbit anti-Phospho-AMPK alpha 1 (S496) antibody (HA750300).
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (HA750300, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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