Anti-phos-AMPKα (Thr172) Antibody, clone 5J23 , ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1213 - recombinant, expressed in HEK 293 cells
Synonym: 5′-AMP-activated protein kinase catalytic subunit alpha-2; ACACA kinase; AMPK subunit alpha-2; Acetyl-CoA carboxylase kinase; EC: 2.7.11.1; HMGCR kinase; Hydroxymethylglutaryl-
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human skin (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1213, Anti-phospho-AMPK (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/nuclear staining was observed in epidermal cells of human skin sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human skin (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1213, Anti-phospho-AMPK (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/nuclear staining was observed in epidermal cells of human skin sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human skin (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1213, Anti-phospho-AMPK (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/nuclear staining was observed in epidermal cells of human skin sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human skin (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1213, Anti-phospho-AMPK (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human skin (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1213, Anti-phospho-AMPK (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Negative control (A) and human skin (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1213, Anti-phospho-AMPK (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HUVEC cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HUVEC cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HUVEC cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1213, Anti-phospho-AMPKα (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitotic cell marker.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from Untreated MCF7 cells (Lane 1) and MCF7 cells treated with Oligomycin (0.5 μM, 30 min) (Lane 2) were probed with Cat. No. ZRB1213, Anti-phospho-AMPK (Thr172), clone 5J23 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Phospho-AMPK (Thr172) (~62 kDa).
Peptide Inhibition Enhanced Validation - Recombinant Antibody Technology Preincubation of a 1:1000 dilution from a representative lot with phosphor-AMPK (Thr172) immunogen peptide abolished target band detection in lysate from MCF7 cells treated with Oligomycin (0.5 μM, 30 min), but not in the corresponding nonphosphopeptide. Peptide block was as follows: Lane 1: Untreated Lane 2: Treated phospho-AMPK (Thr172) peptide added Lane 3: Unmodified AMPK peptide added
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit (recombinant) |
| clone | 5J23, recombinant monoclonal |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 62.32 kDa |
| observed mol wt ~62 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, mouse |
| storage temp. | 2-8°C |
| technique(s) | immunocytochemistry: suitable |
| immunohistochemistry: suitable | |
| inhibition assay: suitable (peptide) | |
| western blot: suitable | |
| UniProt accession no. | P54646  |
| Application: | Anti-phospho-AMPK (Thr172), clone 5J23 ZooMAb, Cat. No. ZRB1213, is a specific recombinant rabbit monoclonal antibody that detects AMPK alpha phosphorylated on threonine 172 and has been tested for use in Immunocytochemistry, Immunohistochemistry, Peptide Inhibition Assay, and Western Blotting. |
| Application: | Peptide Inhibition Analysis: Preincubation of a 1:1000 dilution from a representative lot with phosphor-AMPK (Thr172) immunogen peptide abolished target band detection in lysate from MCF7 cells treated with Oligomycin (0.5 uM, 30 min), but not in the corresponding nonphosphopeptide. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected phospho-AMPK (Thr172) in human skin and human testis tissue sections. Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected phospho-AMPK (Thr172) in HeLa, A431, HUVEC, and NIH 3T3 cells. Note: Actual optimal working dilutions must be determined by end user as specimens and experimental conditions may vary. |
| Disclaimer: | 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. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 10 amino acids surrounding phosphothreonine 172 from human AMPK alpha 2. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 5J23 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects AMPK alpha2. It targets an epitope within 10 amino acids surrounding phosphothreonine 172. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | 5′-AMP-activated protein kinase catalytic subunit alpha-2 (UniProt: P54646; also known as EC: 2.7.11.1, AMPK subunit alpha-2, Acetyl-CoA carboxylase kinase, ACACA kinase, Hydroxymethylglutaryl-CoA reductase kinase, HMGCR kinase) is encoded by the PRKAA2 (also known as AMPK, AMPK2) gene (Gene ID: 5563) in human. AMPK is a heterotrimer composed of an alpha catalytic subunit (PRKAA1 or PRKAA2), a beta (PRKAB1 or PRKAB2) and a gamma non-catalytic subunit (PRKAG1, PRKAG2 or PRKAG3). It also contains an autoinhibitory sequence (aa 291-376) that shows some sequence similarity with the ubiquitin-associated domains and it represses kinase activity. Its protein kinase domains is localized to amino acids 16-268. AMPK is 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, including inhibition of protein, carbohydrate and lipid biosynthesis, and reduces cell growth and proliferation. AMPK acts via direct phosphorylation of metabolic enzymes, and by longer-term effects via phosphorylation of transcription regulators. AMPK is shown to regulate insulin-signaling and glycolysis by phosphorylating IRS1, PFKFB2 and PFKFB3 and it stimulates glucose uptake in muscle by increasing the translocation of the glucose transporter GLUT4 to the plasma membrane. In the liver it acts as a key regulator of glucose homeostasis by phosphorylating CRTC2/TORC2, leading to CRTC2/TORC2 sequestration in the cytoplasm. AMPK is activated by phosphorylation on threonine 172 by STK11/LKB1 or by CAMKK2 triggered by increase in intracellular calcium. Binding of AMP to non-catalytic gamma subunit results in allosteric activation, inducing phosphorylation on this site. AMP-binding to gamma subunit also sustains activity by preventing dephosphorylation of threonine 172. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |