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Phospho-AMPK alpha 1 (S496) Recombinant Rabbit Monoclonal Antibody [SD0810]

Usd: 385

Specification

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Catalog# ET1612-72

Phospho-AMPK alpha 1 (S496) Recombinant Rabbit Monoclonal Antibody [SD0810]

Application

  • WB

  • IP

  • IHC-Fr

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-AMPK alpha 1 (S496) Recombinant Rabbit Monoclonal Antibody [SD0810]

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

RRID

AB_3070148

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:1,000

  • IP

  • Use at an assay dependent concentration.

  • IHC-Fr

  • 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.

UNIPROT

SwissProt: Q13131 Human

SwissProt: Q5EG47 Mouse

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

Expand

Images

  • Western blot analysis of Phospho-AMPK alpha 1 (S496) on different lysates with Rabbit anti-Phospho-AMPK alpha 1 (S496) antibody (<a href="/products/ET1612-72" style="font-weight: bold;text-decoration: underline;">ET1612-72</a>) at 1/5,000 dilution.<br /><br />Lane 1: SiHa cell lysate<br />Lane 2: HUVEC cell lysate<br />Lane 3: K-562 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 64 kDa<br />Observed band size: 64 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1612-72" style="font-weight: bold;text-decoration: underline;">ET1612-72</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Phospho-AMPK alpha 1 (S496) on different lysates with Rabbit anti-Phospho-AMPK alpha 1 (S496) antibody (ET1612-72) 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 (ET1612-72) 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-AMPK alpha 1(S496) on HUVEC cell lysates.<br /><br />Lane 1: HUVEC cells, whole cell lysate, 10ug/lane<br />Lane 2/3: HUVEC cells treated with UV(2 hours recovery), whole cell lysates, 10ug/lane<br />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<br /><br />All lanes :<br />Anti-Phospho-AMPK alpha 1(S496) antibody (<a href="/products/ET1612-72" style="font-weight: bold;text-decoration: underline;">ET1612-72</a>) at 1:500 dilution. Anti-GAPDH antibody (<a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/200,000 dilution.<br /><br />Predicted band size:64 kDa<br />Observed band size:64 kDa<br /><br />Blocking and diluting buffer: 5% BSA.<br /><br />Exposure time: 2 minutes 14 seconds

    ☑ 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 (ET1612-72) 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 (<a href="/products/ET1612-72" style="font-weight: bold;text-decoration: underline;">ET1612-72</a>).<br /><br />The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ET1612-72" style="font-weight: bold;text-decoration: underline;">ET1612-72</a>, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor&reg; 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 hippocampus tissue labeling Phospho-AMPK alpha 1 (S496) with Rabbit anti-Phospho-AMPK alpha 1 (S496) antibody (ET1612-72).

    The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1612-72, 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 (<a href="/products/ET1612-72" style="font-weight: bold;text-decoration: underline;">ET1612-72</a>).<br /><br />The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ET1612-72" style="font-weight: bold;text-decoration: underline;">ET1612-72</a>, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor&reg; 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 (ET1612-72).

    The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1612-72, 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"

Citation

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