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Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) Recombinant Rabbit Monoclonal Antibody [PSH14-13]

Usd: 385

Specification

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Catalog# HA723603

Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) Recombinant Rabbit Monoclonal Antibody [PSH14-13]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

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 (T183) +AMPK alpha 2 (T172) Recombinant Rabbit Monoclonal Antibody [PSH14-13]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phosphopeptide corresponding to residues surrounding Thr183 of human AMPK alpha 1.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 64 kDa

Positive Control

NCI-H1299 cell lysate, NCI-H1299 treated with 0.5μM Oligomycin for 15 minutes cell lysate, SH-SY5Y treated with 0.5μM Oligomycin for 30 minutes cell lysate, C2C12 treated with 1mM AICAR for 30 minutes cell lysate, C6 cell lysate, C6 treated with 1μM Oligomycin for 30 minutes cell lysate, NCI-H1299 treated with 0.5μM Oligomycin for 15 minutes cell lysate then the membrane treated with λpp for 1 hour, human bain tissue untreated / treated, mouse bain tissue untreated / treated, rat bain tissue untreated / treated.

Conjugation

unconjugated

Clone Number

PSH14-13

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P IF-Cell
human
Tested Tested
Tested Tested
Not Recommended
mouse
Tested Tested
Tested Tested
 
rat
Tested Tested
Tested Tested
 

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:200-1:1,000

Target

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

Background References

1. Albayrak G et al. Memantine shifts cancer cell metabolism via AMPK1/2 mediated energetic switch in A549 lung cancer cells. EXCLI J. 2021 Feb

2. Lin FC et al.Protective Effects of Kirenol against Lipopolysaccharide-Induced Acute Lung Injury through the Modulation of the Proinflammatory NFkappaB Pathway and the AMPK2-/Nrf2-Mediated HO-1/AOE Pathway. Antioxidants (Basel). 2021 Jan

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q13131 Human

SwissProt: P54646 Human

SwissProt: Q5EG47 Mouse

SwissProt: Q8BRK8 Mouse

SwissProt: P54645 Rat

SwissProt: Q09137 Rat

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) on different lysates with Rabbit anti-Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) antibody (<a href="/products/HA723603" style="font-weight: bold;text-decoration: underline;">HA723603</a>) at 1/2,000 dilution.<br /><br />Lane 1: NCI-H1299 cell lysate<br />Lane 2: NCI-H1299 treated with 0.5μM Oligomycin for 15 minutes cell lysate<br />Lane 3: SH-SY5Y cell lysate<br />Lane 4: SH-SY5Y treated with 0.5μM Oligomycin for 30 minutes cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: C2C12 treated with 1mM AICAR for 30 minutes cell lysate<br />Lane 7: C6 cell lysate<br />Lane 8: C6 treated with 1μM Oligomycin for 30 minutes cell lysate<br />Lane 9: NCI-H1299 treated with 0.5μM Oligomycin for 15 minutes cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 64 kDa<br />Observed band size: 64 kDa<br /><br />Exposure time: 59 seconds; ECL: K1801;<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/HA723603" style="font-weight: bold;text-decoration: underline;">HA723603</a>) at 1/2,000 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.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) on different lysates with Rabbit anti-Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) antibody (HA723603) at 1/2,000 dilution.

    Lane 1: NCI-H1299 cell lysate
    Lane 2: NCI-H1299 treated with 0.5μM Oligomycin for 15 minutes cell lysate
    Lane 3: SH-SY5Y cell lysate
    Lane 4: SH-SY5Y treated with 0.5μM Oligomycin for 30 minutes cell lysate
    Lane 5: C2C12 cell lysate
    Lane 6: C2C12 treated with 1mM AICAR for 30 minutes cell lysate
    Lane 7: C6 cell lysate
    Lane 8: C6 treated with 1μM Oligomycin for 30 minutes cell lysate
    Lane 9: NCI-H1299 treated with 0.5μM Oligomycin for 15 minutes cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 64 kDa
    Observed band size: 64 kDa

    Exposure time: 59 seconds; ECL: K1801;

    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 (HA723603) at 1/2,000 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 />Immunohistochemical analysis of paraffin-embedded human bain tissue untreated / treated with λpp with Rabbit anti-Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) antibody (<a href="/products/HA723603" style="font-weight: bold;text-decoration: underline;">HA723603</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723603" style="font-weight: bold;text-decoration: underline;">HA723603</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    ☑ Cell treatment (CT)

    Immunohistochemical analysis of paraffin-embedded human bain tissue untreated / treated with λpp with Rabbit anti-Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) antibody (HA723603) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723603) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunohistochemical analysis of paraffin-embedded mouse bain tissue untreated / treated with λpp with Rabbit anti-Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) antibody (<a href="/products/HA723603" style="font-weight: bold;text-decoration: underline;">HA723603</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723603" style="font-weight: bold;text-decoration: underline;">HA723603</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    ☑ Cell treatment (CT)

    Immunohistochemical analysis of paraffin-embedded mouse bain tissue untreated / treated with λpp with Rabbit anti-Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) antibody (HA723603) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723603) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunohistochemical analysis of paraffin-embedded rat bain tissue untreated / treated with λpp with Rabbit anti-Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) antibody (<a href="/products/HA723603" style="font-weight: bold;text-decoration: underline;">HA723603</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723603" style="font-weight: bold;text-decoration: underline;">HA723603</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    ☑ Cell treatment (CT)

    Immunohistochemical analysis of paraffin-embedded rat bain tissue untreated / treated with λpp with Rabbit anti-Phospho-AMPK alpha 1 (T183) +AMPK alpha 2 (T172) antibody (HA723603) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723603) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Citation

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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