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AMPK alpha 1 Overexpression Lysate

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Western Blot: AMPK alpha 1 Overexpression Lysate (Adult Normal) [NBP2-10421] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for AMPK alpha 1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

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AMPK alpha 1 Overexpression Lysate Summary

Description

AMPK alpha 1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC218572

Accession#: NM_006251

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Gene
PRKAA1

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW
63.8 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for AMPK alpha 1 Overexpression Lysate

  • AMP -activate kinase alpha 1 subunit
  • AMP-activated protein kinase, catalytic, alpha-1,5'-AMP-activated protein kinase catalytic subunit alpha-1
  • AMPK alpha 1
  • AMPK alpha 1,5'-AMP-activated protein kinase, catalytic alpha-1 chain
  • AMPK subunit alpha-1
  • AMPK
  • AMPK1
  • AMPKa1
  • EC 2.7.11
  • EC 2.7.11.1
  • MGC33776
  • MGC57364
  • PRKAA1
  • protein kinase, AMP-activated, alpha 1 catalytic subunit

Background

The 5'-AMP-activated protein kinase (AMPK), a member of the SNF1 (sucrose non-fermentor) kinase family (1) AMPK is a heterotrimeric protein comprising alpha (63 kDa), beta (38 kDa) and gamma (38 kDa) subunits. The alpha subunit is the catalytic subunit (which requires phosphorylation to be active) (2-3). Beta and gamma are non-catalytic subunits, although they have been found to interact with the active subunit in liver. AMPK regulates fatty acid and sterol synthesis by phosphorylation of acetyl-CoA as well as cholesterol synthesis via phosphorylation and inactivation of hydroxymethylglutaryl-CoA reductase (4). AMPK also appears to play a role in protecting cells from stresses that cause ATP depletion by switching off ATP-consuming biosynthetic pathways

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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AMP-activated protein kinase (AMPK) is best known as a sensor of oxidative stress.  AMPK is activated by increased intracellular AMP levels, which are a result of alterations in cellular metabolism from causes such as hypoxia, changes in ATP, sene...  Read full blog post.

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Bioinformatics

Gene Symbol PRKAA1