PRODUCT CODE: ET1612-72

Recombinant Phospho AMPK alpha 1 (S496) Monoclonal Antibody (ET1612-72)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IP

REACTIVITY

  • Human

Western blot analysis of Phospho-AMPK alpha 1 (S496) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-72, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: 293T cell lysate<br />
 Lane 2: SiHa cell lysate
  • Western blot analysis of Phospho-AMPK alpha 1 (S496) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-72, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: 293T cell lysate<br />
 Lane 2: SiHa cell lysate
  • ICC staining of Phospho-AMPK alpha 1 (S496) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-72, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-AMPK alpha 1 (S496) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-72, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-AMPK alpha 1 (S496) in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-72, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Western blot analysis of Phospho-AMPK alpha 1 (S496) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-72, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: 293T cell lysate
Lane 2: SiHa cell lysate

Applications

  • WB

  • ICC

  • IF

  • IP

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho AMPK alpha 1 (S496) Monoclonal Antibody (ET1612-72)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding ser496 of human ampk alpha 1.

Host

Rabbit

Modification

Phospho

Modification Site

S496

Positive Control

293T cell lysate, SiHa cell lysate, Hela, A549, 293T.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SD0810

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

64 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC/IF

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Phospho-AMPK alpha 1 (S496)

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; AL024255 antibody; AMP -activate kinase alpha 1 subunit antibody; AMP-activated protein kinase, catalytic, alpha -1 antibody; AMPK 1 antibody; AMPK alpha 1 antibody; AMPK alpha 1 chain antibody; AMPK antibody; AMPK subunit alpha-1 antibody; AMPK1 antibody; AMPKa1 antibody; AMPKalpha1 antibody; C130083N04Rik antibody; cb116 antibody; EC 2.7.11.1 antibody; HMG CoA reductase kinase antibody; HMGCR kinase antibody; hormone sensitive lipase kinase antibody; Hydroxymethylglutaryl CoA reductase kinase antibody; im:7154392 antibody; kinase AMPK alpha1 antibody; MGC33776 antibody; MGC57364 antibody; OTTHUMP00000161795 antibody; OTTHUMP00000161796 antibody; PRKAA 1 antibody; PRKAA1 antibody; Protein kinase AMP activated alpha 1 catalytic subunit antibody; SNF1-like protein AMPK antibody; SNF1A antibody; Tau protein kinase PRKAA1 antibody; wu:fa94c10 antibody

SEQUENCE SIMILARITIES

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.

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.