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Western Blot Analysis
Non-stimulated A431cell lysate was resolved by electrophoresis, transferred to nitrocellulose and
probed with anti-AM...
Western Blot Analysis
A rat liver preparation was resolved by
electrophoresis, transferred to nitrocellulose and probed with anti-AMPKα2 (2...
)
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AMPK Pathway Explorer Antibody MiniPack
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M, R | WB | Rabbit | Purified | Polyclonal Antibody |
Description:
AMPK Pathway Explorer Antibody MiniPack
Trade Name:
Upstate (Millipore)
Qty/Pk:
3 vials/Pk
Background Information:
Pathway Explorer Antibody MiniPack:
Each Pathway Explorer Antibody Minipack contains three related antibodies as part of a signaling cascade or a combination of total and phosphorylated forms of key signaling targets. Each of the three antibodies are 30% the original pack size. Full size versions of each of the Pathway Explorer antibodies are available for sale individually under the same catalog number with the removal of “SP” off of each one (e.g. 05-591SP can be ordered as 05-591).
AMPK:
AMP-activated protein kinase (AMPK) is a metabolic and stress-sensing kinase that regulates homeostasis, and a key target for treating Type 2 diabetes and obesity. AMPK exists as a heterotrimeric complex comprised of catalytic alpha subunits (62 kDa) and non-catalytic beta and gamma subunits. Alpha subunit has at least two isoforms (alpha 1 and alpha 2) which differ in their subcellular localization and AMP-dependence. AMPK is phosphorylated by upstream kinases, AMPK Kinase (AMPKK) and LKB1 which results in its activation. Active AMPK regulates metabolism by phosphorylating rate-limiting enzymes in metabolic pathways and controlling gene expression. Phosphorylation of threonine 172 in the activation loop of the alpha subunit is a key determinant of AMPK activity.
*See full size versions for corresponding references.
Each Pathway Explorer Antibody Minipack contains three related antibodies as part of a signaling cascade or a combination of total and phosphorylated forms of key signaling targets. Each of the three antibodies are 30% the original pack size. Full size versions of each of the Pathway Explorer antibodies are available for sale individually under the same catalog number with the removal of “SP” off of each one (e.g. 05-591SP can be ordered as 05-591).
AMPK:
AMP-activated protein kinase (AMPK) is a metabolic and stress-sensing kinase that regulates homeostasis, and a key target for treating Type 2 diabetes and obesity. AMPK exists as a heterotrimeric complex comprised of catalytic alpha subunits (62 kDa) and non-catalytic beta and gamma subunits. Alpha subunit has at least two isoforms (alpha 1 and alpha 2) which differ in their subcellular localization and AMP-dependence. AMPK is phosphorylated by upstream kinases, AMPK Kinase (AMPKK) and LKB1 which results in its activation. Active AMPK regulates metabolism by phosphorylating rate-limiting enzymes in metabolic pathways and controlling gene expression. Phosphorylation of threonine 172 in the activation loop of the alpha subunit is a key determinant of AMPK activity.
*See full size versions for corresponding references.
Key Applications:
Western Blotting
Species Reactivity:
- Human
- Mouse
- Rat
Species Reactivity Note:
07-681SP:Human and rat. Expected to cross-react with mouse based on 100% sequence homology.
07-363SP: Human and rat.
07-350SP: Human, rat, and mouse.
07-363SP: Human and rat.
07-350SP: Human, rat, and mouse.
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Entrez Gene Summary:
AMPK: The protein encoded by this gene belongs to the ser/thr protein kinase family. It is the catalytic subunit of the 5'-prime-AMP-activated protein kinase (AMPK). AMPK is a cellular energy sensor conserved in all eukaryotic cells. The kinase activity of AMPK is activated by the stimuli that increase the cellular AMP/ATP ratio. AMPK regulates the activities of a number of key metabolic enzymes through phosphorylation. It protects cells from stresses that cause ATP depletion by switching off ATP-consuming biosynthetic pathways. Alternatively spliced transcript variants encoding distinct isoforms have been observed.
UniProt Summary:
FUNCTION: Responsible for the regulation of fatty acid synthesis by phosphorylation of acetyl-CoA carboxylase. It also regulates cholesterol synthesis via phosphorylation and inactivation of hormone-sensitive lipase and hydroxymethylglutaryl-CoA reductase. Appears to act as a metabolic stress-sensing protein kinase switching off biosynthetic pathways when cellular ATP levels are depleted and when 5'-AMP rises in response to fuel limitation and/or hypoxia. This is a catalytic subunit. Ref.8 Ref.9
Catalytic activity ATP + a protein = ADP + a phosphoprotein.
Cofactor Magnesium.
ENZYME REGULATION: Binding of AMP results in allosteric activation, inducing phosphorylation on Thr-174 by STK11 in complex with STE20-related adapter-alpha (STRAD alpha) pseudo kinase and CAB39. Also activated by phosphorylation by CAMKK2 triggered by a rise in intracellular calcium ions, without detectable changes in the AMP/ATP ratio. Ref.8 Ref.9
SUBUNIT STRUCTURE: Heterotrimer of an alpha catalytic subunit, a beta and a gamma non-catalytic subunits. Interacts with FNIP1 and FNIP2. Ref.14 Ref.15
SEQUENCE SIMILARITY: Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
Contains 1 protein kinase domain.
SEQUENCE CAUTION: The sequence AAH48980.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened
Catalytic activity ATP + a protein = ADP + a phosphoprotein.
Cofactor Magnesium.
ENZYME REGULATION: Binding of AMP results in allosteric activation, inducing phosphorylation on Thr-174 by STK11 in complex with STE20-related adapter-alpha (STRAD alpha) pseudo kinase and CAB39. Also activated by phosphorylation by CAMKK2 triggered by a rise in intracellular calcium ions, without detectable changes in the AMP/ATP ratio. Ref.8 Ref.9
SUBUNIT STRUCTURE: Heterotrimer of an alpha catalytic subunit, a beta and a gamma non-catalytic subunits. Interacts with FNIP1 and FNIP2. Ref.14 Ref.15
SEQUENCE SIMILARITY: Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
Contains 1 protein kinase domain.
SEQUENCE CAUTION: The sequence AAH48980.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened
Molecular Weight:
~63 kDa;
Brand Family:
Upstate
Control:
- For 07-681SP use HEK293 cells or rat liver AMPK.
- For 07-363SP use Rat liver preparation.
- For 07-350SP use A431cell lysate, or lysate of HeLa cells treated with heat shock.
Presentation:
3 individual tubes containing either Anti-AMPK α1; Anti-AMPKα2; or Anti-phospho-AMPKα (Thr172)
Storage Conditions:
Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.
Modifications:
Phosphorylation
Gene Symbol:
AMPK
Cross Reactivity:
Varies. See individual Data Sheets
Product Name:
AMPK Pathway Explorer Antibody MiniPack
Alternate Names:
AMP-Activated Protein Kinase
Concentration:
1 mg/mL
Antibody Type:
Polyclonal Antibody
Specificity:
07-681SP: AMPKα phosphorylated on Thr172.
07-363SP: AMP-Activated Protein Kinase alpha 2 (AMPKα2).
07-350SP: AMPKα1
07-363SP: AMP-Activated Protein Kinase alpha 2 (AMPKα2).
07-350SP: AMPKα1
Quality Assurance:
07-681SP:Routinely evaluated by immunoblot on RIPA lysates from overnight starved HEK293 cells and partially purified rat liver AMPK.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected phospho-AMPK α in RIPA lysates from overnight serum starved HEK293 cells and partially purified rat liver AMPK. Signal was reduced after treatment of lysate with λ-phosphatase.
07-363SP: Routinely evaluated by immunoblot in a rat liver preparation.
Western Blot Analysis:
1-2 μg/mL of this lot detected AMPK α2 in a rat liver preparation.
07-350SP: Routinely evaluated by immunoblot in RIPA lysates from non-stimulated A431 cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected AMPK α1 in RIPA lysates from non-stimulated A431 cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected phospho-AMPK α in RIPA lysates from overnight serum starved HEK293 cells and partially purified rat liver AMPK. Signal was reduced after treatment of lysate with λ-phosphatase.
07-363SP: Routinely evaluated by immunoblot in a rat liver preparation.
Western Blot Analysis:
1-2 μg/mL of this lot detected AMPK α2 in a rat liver preparation.
07-350SP: Routinely evaluated by immunoblot in RIPA lysates from non-stimulated A431 cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected AMPK α1 in RIPA lysates from non-stimulated A431 cells.
Immunogen:
Varies. See individual Data Sheets
Components:
- 07-350SP Anti-AMPK α1; 60 µg
- 07-363SP Anti-AMPKα2; 60 µg
- 07-681SP Anti-phospho-AMPKα (Thr172); 60 µg
Format:
Purified
UniProt Number:
Host:
Rabbit
Properties:
Each vial is 30% the size of the parent catalog number
Epitope:
07-350SP and 07-363SP: Internal
Entrez Gene Number:
Purification Method:
Protein A purfied