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  • Product Name
    Phospho-AMPK alpha (Thr172) Antibody
  • Catalog No.
    AF3423
  • RRID
    AB_2834865
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity
    Human, Mouse, Rat
  • Prediction
    Pig(100%), Zebrafish(100%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
  • UniProt
  • Mol.Wt
    62kD;
    64kDa,62kDa(Calculated).
  • Concentration
    1mg/ml
  • Browse similar products>>

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Product Information

Alternative Names:Expand▼

5 AMP activated protein kinase alpha 1catalytic subunit; 5 AMP activated protein kinase catalytic alpha 1 chain; 5' AMP activated protein kinase catalytic subunit alpha 1; 5'-AMP-activated protein kinase catalytic subunit alpha-1; AAPK1; AAPK1_HUMAN; ACACA kinase; acetyl CoA carboxylase kinase; AI194361; AI450832; AL024255; AMP -activate kinase alpha 1 subunit; AMP-activated protein kinase, catalytic, alpha -1; AMPK 1; AMPK alpha 1; AMPK alpha 1 chain; AMPK; AMPK subunit alpha-1; AMPK1; AMPKa1; AMPKalpha1; C130083N04Rik; cb116; EC 2.7.11.1; HMG CoA reductase kinase; HMGCR kinase; hormone sensitive lipase kinase; Hydroxymethylglutaryl CoA reductase kinase; im:7154392; kinase AMPK alpha1; MGC33776; MGC57364; OTTHUMP00000161795; OTTHUMP00000161796; PRKAA 1; PRKAA1; Protein kinase AMP activated alpha 1 catalytic subunit; SNF1-like protein AMPK; SNF1A; Tau protein kinase PRKAA1; wu:fa94c10; 5'-AMP-activated protein kinase catalytic subunit alpha-2; AAPK2_HUMAN; ACACA kinase; Acetyl-CoA carboxylase kinase; AMPK alpha 2 chain; AMPK subunit alpha-2; AMPK2; AMPKa2; AMPKalpha2; HMGCR kinase; Hydroxymethylglutaryl-CoA reductase kinase; PRKAA; PRKAA2; Protein kinase AMP activated alpha 2 catalytic subunit; Protein kinase AMP activated catalytic subunit alpha 2;

Applications:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-500, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Pig(100%), Zebrafish(100%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Specificity:

Phospho-AMPK alpha (Thr172) Antibody detects endogenous levels of AMPK alpha only when phosphorylated at Threonine 172.

RRID:

AB_2834865
Please cite this product as: Affinity Biosciences Cat# AF3423, RRID:AB_2834865.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

Immunogen Information

Immunogen:

A synthesized peptide derived from human AMPK alpha around the phosphorylation site of Thr172.

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

PRKAA1,PRKAA2

Molecular Weight:

Observed Mol.Wt.: 62kD.
Predicted Mol.Wt.: 64kDa,62kDa(Calculated)..

Description:

AMPKA2 a protein kinase of the CAMKL family. The holoenzyme consists of a catalytic subunit (alpha) and two regulatory subunits (beta, gamma).

Sequence:
MRRLSSWRKMATAEKQKHDGRVKIGHYILGDTLGVGTFGKVKVGKHELTGHKVAVKILNRQKIRSLDVVGKIRREIQNLKLFRHPHIIKLYQVISTPSDIFMVMEYVSGGELFDYICKNGRLDEKESRRLFQQILSGVDYCHRHMVVHRDLKPENVLLDAHMNAKIADFGLSNMMSDGEFLRTSCGSPNYAAPEVISGRLYAGPEVDIWSSGVILYALLCGTLPFDDDHVPTLFKKICDGIFYTPQYLNPSVISLLKHMLQVDPMKRATIKDIREHEWFKQDLPKYLFPEDPSYSSTMIDDEALKEVCEKFECSEEEVLSCLYNRNHQDPLAVAYHLIIDNRRIMNEAKDFYLATSPPDSFLDDHHLTRPHPERVPFLVAETPRARHTLDELNPQKSKHQGVRKAKWHLGIRSQSRPNDIMAEVCRAIKQLDYEWKVVNPYYLRVRRKNPVTSTYSKMSLQLYQVDSRTYLLDFRSIDDEITEAKSGTATPQRSGSVSNYRSCQRSDSDAEAQGKSSEVSLTSSVTSLDSSPVDLTPRPGSHTIEFFEMCANLIKILAQ

MAEKQKHDGRVKIGHYVLGDTLGVGTFGKVKIGEHQLTGHKVAVKILNRQKIRSLDVVGKIKREIQNLKLFRHPHIIKLYQVISTPTDFFMVMEYVSGGELFDYICKHGRVEEMEARRLFQQILSAVDYCHRHMVVHRDLKPENVLLDAHMNAKIADFGLSNMMSDGEFLRTSCGSPNYAAPEVISGRLYAGPEVDIWSCGVILYALLCGTLPFDDEHVPTLFKKIRGGVFYIPEYLNRSVATLLMHMLQVDPLKRATIKDIREHEWFKQDLPSYLFPEDPSYDANVIDDEAVKEVCEKFECTESEVMNSLYSGDPQDQLAVAYHLIIDNRRIMNQASEFYLASSPPSGSFMDDSAMHIPPGLKPHPERMPPLIADSPKARCPLDALNTTKPKSLAVKKAKWHLGIRSQSKPYDIMAEVYRAMKQLDFEWKVVNAYHLRVRRKNPVTGNYVKMSLQLYLVDNRSYLLDFKSIDDEVVEQRSGSSTPQRSCSAAGLHRPRSSFDSTTAESHSLSGSLTGSLTGSTLSSVSPRLGSHTMDFFEMCASLITTLAR

Research Background

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. Also acts as a regulator of cellular polarity by remodeling the actin cytoskeleton; probably by indirectly activating myosin. Regulates lipid synthesis by phosphorylating and inactivating lipid metabolic enzymes such as ACACA, ACACB, GYS1, HMGCR and LIPE; regulates fatty acid and cholesterol synthesis by phosphorylating acetyl-CoA carboxylase (ACACA and ACACB) and hormone-sensitive lipase (LIPE) enzymes, respectively. Regulates insulin-signaling and glycolysis by phosphorylating IRS1, PFKFB2 and PFKFB3. AMPK stimulates glucose uptake in muscle by increasing the translocation of the glucose transporter SLC2A4/GLUT4 to the plasma membrane, possibly by mediating phosphorylation of TBC1D4/AS160. Regulates transcription and chromatin structure by phosphorylating transcription regulators involved in energy metabolism such as CRTC2/TORC2, FOXO3, histone H2B, HDAC5, MEF2C, MLXIPL/ChREBP, EP300, HNF4A, p53/TP53, SREBF1, SREBF2 and PPARGC1A. Acts as a key regulator of glucose homeostasis in liver by phosphorylating CRTC2/TORC2, leading to CRTC2/TORC2 sequestration in the cytoplasm. In response to stress, phosphorylates 'Ser-36' of histone H2B (H2BS36ph), leading to promote transcription. Acts as a key regulator of cell growth and proliferation by phosphorylating TSC2, RPTOR and ATG1/ULK1: in response to nutrient limitation, negatively regulates the mTORC1 complex by phosphorylating RPTOR component of the mTORC1 complex and by phosphorylating and activating TSC2. In response to nutrient limitation, promotes autophagy by phosphorylating and activating ATG1/ULK1. In that process also activates WDR45 (PubMed:28561066). In response to nutrient limitation, phosphorylates transcription factor FOXO3 promoting FOXO3 mitochondrial import (By similarity). AMPK also acts as a regulator of circadian rhythm by mediating phosphorylation of CRY1, leading to destabilize it. May regulate the Wnt signaling pathway by phosphorylating CTNNB1, leading to stabilize it. Also has tau-protein kinase activity: in response to amyloid beta A4 protein (APP) exposure, activated by CAMKK2, leading to phosphorylation of MAPT/TAU; however the relevance of such data remains unclear in vivo. Also phosphorylates CFTR, EEF2K, KLC1, NOS3 and SLC12A1.

Post-translational Modifications:

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:

Cytoplasm. Nucleus.
Note: In response to stress, recruited by p53/TP53 to specific promoters.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Subunit Structure:

AMPK is a heterotrimer of an alpha catalytic subunit (PRKAA1 or PRKAA2), a beta (PRKAB1 or PRKAB2) and a gamma non-catalytic subunits (PRKAG1, PRKAG2 or PRKAG3). Interacts with FNIP1 and FNIP2.

Similarity:

The AIS (autoinhibitory sequence) region shows some sequence similarity with the ubiquitin-associated domains and represses kinase activity.

Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.

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. Also acts as a regulator of cellular polarity by remodeling the actin cytoskeleton; probably by indirectly activating myosin. Regulates lipid synthesis by phosphorylating and inactivating lipid metabolic enzymes such as ACACA, ACACB, GYS1, HMGCR and LIPE; regulates fatty acid and cholesterol synthesis by phosphorylating acetyl-CoA carboxylase (ACACA and ACACB) and hormone-sensitive lipase (LIPE) enzymes, respectively. Regulates insulin-signaling and glycolysis by phosphorylating IRS1, PFKFB2 and PFKFB3. Involved in insulin receptor/INSR internalization (PubMed:25687571). AMPK stimulates glucose uptake in muscle by increasing the translocation of the glucose transporter SLC2A4/GLUT4 to the plasma membrane, possibly by mediating phosphorylation of TBC1D4/AS160. Regulates transcription and chromatin structure by phosphorylating transcription regulators involved in energy metabolism such as CRTC2/TORC2, FOXO3, histone H2B, HDAC5, MEF2C, MLXIPL/ChREBP, EP300, HNF4A, p53/TP53, SREBF1, SREBF2 and PPARGC1A. Acts as a key regulator of glucose homeostasis in liver by phosphorylating CRTC2/TORC2, leading to CRTC2/TORC2 sequestration in the cytoplasm. In response to stress, phosphorylates 'Ser-36' of histone H2B (H2BS36ph), leading to promote transcription. Acts as a key regulator of cell growth and proliferation by phosphorylating TSC2, RPTOR and ATG1/ULK1: in response to nutrient limitation, negatively regulates the mTORC1 complex by phosphorylating RPTOR component of the mTORC1 complex and by phosphorylating and activating TSC2. In response to nutrient limitation, promotes autophagy by phosphorylating and activating ATG1/ULK1. In that process also activates WDR45 (PubMed:28561066). AMPK also acts as a regulator of circadian rhythm by mediating phosphorylation of CRY1, leading to destabilize it. May regulate the Wnt signaling pathway by phosphorylating CTNNB1, leading to stabilize it. Also phosphorylates CFTR, EEF2K, KLC1, NOS3 and SLC12A1. Plays an important role in the differential regulation of pro-autophagy (composed of PIK3C3, BECN1, PIK3R4 and UVRAG or ATG14) and non-autophagy (composed of PIK3C3, BECN1 and PIK3R4) complexes, in response to glucose starvation. Can inhibit the non-autophagy complex by phosphorylating PIK3C3 and can activate the pro-autophagy complex by phosphorylating BECN1 (By similarity).

Post-translational Modifications:

Ubiquitinated.

Phosphorylated at Thr-172 by STK11/LKB1 in complex with STE20-related adapter-alpha (STRADA) pseudo kinase and CAB39. Also phosphorylated at Thr-172 by CAMKK2; triggered by a rise in intracellular calcium ions, without detectable changes in the AMP/ATP ratio. CAMKK1 can also phosphorylate Thr-172, but at much lower level. Dephosphorylated by protein phosphatase 2A and 2C (PP2A and PP2C). Phosphorylated by ULK1; leading to negatively regulate AMPK activity and suggesting the existence of a regulatory feedback loop between ULK1 and AMPK. Dephosphorylated by PPM1A and PPM1B at Thr-172 (mediated by STK11/LKB1).

Subcellular Location:

Cytoplasm. Nucleus.
Note: In response to stress, recruited by p53/TP53 to specific promoters.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Subunit Structure:

AMPK is a heterotrimer of an alpha catalytic subunit (PRKAA1 or PRKAA2), a beta (PRKAB1 or PRKAB2) and a gamma non-catalytic subunits (PRKAG1, PRKAG2 or PRKAG3). Interacts with FNIP1 and FNIP2. Associates with internalized insulin receptor/INSR complexes on Golgi/endosomal membranes; PRKAA2/AMPK2 together with ATIC and HACD3/PTPLAD1 is proposed to be part of a signaling network regulating INSR autophosphorylation and endocytosis (PubMed:25687571).

Similarity:

The AIS (autoinhibitory sequence) region shows some sequence similarity with the ubiquitin-associated domains and represses kinase activity.

Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.

Research Fields

Research Fields:

· Cellular Processes > Cellular community - eukaryotes > Tight junction.(View pathway)
· Cellular Processes > Transport and catabolism > Autophagy - animal.(View pathway)
· Environmental Information Processing > Signal transduction > FoxO signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > mTOR signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Apelin signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > AMPK signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.(View pathway)
· Human Diseases > Cardiovascular diseases > Hypertrophic cardiomyopathy (HCM).
· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.
· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).
· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.
· Organismal Systems > Endocrine system > Insulin signaling pathway.(View pathway)
· Organismal Systems > Endocrine system > Glucagon signaling pathway.
· Organismal Systems > Environmental adaptation > Circadian rhythm.(View pathway)
· Organismal Systems > Endocrine system > Oxytocin signaling pathway.
· Organismal Systems > Aging > Longevity regulating pathway.(View pathway)
· Organismal Systems > Aging > Longevity regulating pathway - multiple species.(View pathway)

Reference Citations:

1). Zhang F et al. Anti-osteoporosis activity of Sanguinarine in preosteoblast MC3T3-E1 cells and an ovariectomized rat model. J Cell Physiol 2018 Jun;233(6):4626-4633 (PubMed: 28926099) [IF=5.546]

Application: WB    Species:mouse;    Sample:Not available

Figure 3. Compound C reversed the effects of Sanguinarine on the differentiation of MC3T3-E1 cells. MC3T3-E1 cells were pretreated with 10 μM Compound C for 1 h and then exposed to 2 M Sanguinarine (SAN). Mock cells were cultured without any treatment. (A, B) At 48 h after treatment, the levels of AMPK1 and p-AMPK (A), and ALP activity in the cultured medium (B) were analyzed. (C) mRNA levels of Bmp2, Osx and Opg were detected by real-time PCR. (D) Protein levels of Bmp2, Smad1, pSmad1, Osx and Opg were determined by Western blot. ***P <0.001 versus DMSO; +++P <0.001 versus Compound C.


2). Li M et al. Respiratory Syncytial Virus Replication Is Promoted by Autophagy-Mediated Inhibition of Apoptosis. J Virol 2018 Mar 28;92(8) (PubMed: 29386287) [IF=4.501]

3). Li M et al. Respiratory Syncytial Virus Replication Is Promoted by Autophagy-Mediated Inhibition of Apoptosis. J Virol 2018 Mar 28;92(8) (PubMed: 29386287) [IF=4.501]

4). Wang L et al. Inhibition of GSK3β Reduces Ectopic Lipid Accumulation and Induces Autophagy by the AMPK Pathway in Goat Muscle Satellite Cells. Cells 2019 Nov 1;8(11) (PubMed: 31683987) [IF=4.366]

5). Liu S et al. Cerasus humilis cherry polyphenol reduces high-fat diet-induced obesity in C57BL/6 mice by mitigating fat deposition, inflammation, and oxidation. J Agric Food Chem 2020 Mar 30 (PubMed: 32227855) [IF=4.192]

6). Xiao L et al. Hydrogen sulfide improves endothelial dysfunction in hypertension by activating peroxisome proliferator-activated receptor delta/endothelial nitric oxide synthase signaling. J Hypertens 2018 Mar;36(3):651-665 (PubMed: 29084084) [IF=4.171]

7). Zhang X et al. Protective effects of protocatechuic acid on acute lung injury induced by lipopolysaccharide in mice via p38MAPK and NF-κB signal pathways. Int Immunopharmacol 2015 May;26(1):229-36 (PubMed: 25841318) [IF=3.943]

8). Huang X et al. High expressions of LDHA and AMPK as prognostic biomarkers for breast cancer. Breast 2016 Dec;30:39-46 (PubMed: 27598996) [IF=3.754]

Application: WB    Species:human;    Sample:Not available

Fig. 1. LDHA and AMPK were up-regulated synchronously in breast cancer. A. Expression levels of LDHA, AMPK and pAMPK were assessed by Western blot (above) and gray image scanning (below) in eight different breast cancer cell lines, including four NTNBC cell lines and four TNBC cell lines. GAPDH was used as a loading control. B. Expression levels of LDHA and AMPK were determined by qRT-PCR (above). The differences between TNBC and NTNBC cell lines were analyzed (below). GAPDH was used as an internal control. C. Expression levels of LDHA, AMPK and pAMPK were assessed by Western blot (above) and gray image scanning (below) in eight different breast cancer tissues, including four NTNBC tissues and four TNBC tissues. GAPDH was used as a loading control. D. Expression levels of LDHA and AMPK were determined by qRT-PCR (above). The diffe


9). Lu W et al. ApoE deficiency promotes non-alcoholic fatty liver disease in mice via impeding AMPK/mTOR mediated autophagy. Life Sci 2020 Apr 15;252:117601 (PubMed: 32304762) [IF=3.647]

10). Ao LY et al. Therapeutic effects of JLX-001 on ischemic stroke by inducing autophagy via AMPK-ULK1 signaling pathway in rats. Brain Res Bull 2019 Aug 28;153:162-170 (PubMed: 31472184) [IF=3.370]

11). Li X et al. Enhancement of Glucose Metabolism via PGC-1α Participates in the Cardioprotection of Chronic Intermittent Hypobaric Hypoxia. Front Physiol 2016 Jun 8;7:219 (PubMed: 27375497) [IF=3.367]

Application: WB    Species:rat;    Sample:Not available

FIGURE 4 | Effect of CIHH on the protein expression of AMPK, p-AMPK, PDK4 and PGC-1α in left ventricular myocytes before and after I/R. (A) Protein expression of AMPK and p-AMPK; (B) Protein expression of PDK4; (C) Protein expression of PGC-1α. CON, control group; CIHH, chronic intermittent hypobaric hypoxia; Pre, pre-ischemia; I/R, ischemia/reperfusion. Data are expressed as the mean ± SD (n = 6 in each group). *P < 0.05 vs. corresponding CON group.


12). Lin H et al. Rapamycin Supplementation May Ameliorate Erectile Function in Rats With Streptozotocin-Induced Type 1 Diabetes by Inducing Autophagy and Inhibiting Apoptosis, Endothelial Dysfunction, and Corporal Fibrosis. J Sex Med 2018 Sep;15(9):1246-1259 (PubMed: 30224017) [IF=3.293]

13). Yin X et al. The intraperitoneal administration of MOTS-c produces antinociceptive and anti-inflammatory effects through the activation of AMPK pathway in the mouse formalin test. Eur J Pharmacol 2020 Jan 8;870:172909 (PubMed: 31926126) [IF=3.263]

14). Li S et al. Effect of CAPE-pNO2 against type 2 diabetes mellitus via the AMPK/GLUT4/ GSK3β/PPARα pathway in HFD/STZ-induced diabetic mice. Eur J Pharmacol 2019 Mar 15;853:1-10 (PubMed: 30885574) [IF=3.263]

15). Cheng Y et al. Strontium promotes osteogenic differentiation by activating autophagy via the the AMPK/mTOR signaling pathway in MC3T3‑E1 cells. Int J Mol Med 2019 Aug;44(2):652-660 (PubMed: 31173178) [IF=3.098]

16). Cheng Y et al. Strontium promotes osteogenic differentiation by activating autophagy via the the AMPK/mTOR signaling pathway in MC3T3‑E1 cells. Int J Mol Med 2019 Aug;44(2):652-660 (PubMed: 31173178) [IF=3.098]

17). Dong W et al. Zoledronate and high glucose levels influence osteoclast differentiation and bone absorption via the AMPK pathway. Biochem Biophys Res Commun 2018 Oct 12 (PubMed: 30322621)

18). Deng Z et al. Myostatin inhibits eEF2K-eEF2 by regulating AMPK to suppress protein synthesis. Biochem Biophys Res Commun 2017 Dec 9;494(1-2):278-284 (PubMed: 29024627)

Application: WB    Species:mouse;    Sample:Not available

Fig. 4. Myostatin regulated translation elongation through AMP. C2C12 myotubes were treated with various concentration recombinant myostatin (0, 0.01,0.1, 1, 2, 3 µg/ml) for 48 h andthen lysed and cellular extracts were analyzed by Western blot with anti-AMPK(A).


19). Meng Z et al. Hepatitis C virus nonstructural protein 5A perturbs lipid metabolism by modulating AMPK/SREBP-1c signaling. Lipids Health Dis 2019 Nov 4;18(1):191 (PubMed: 31684957)

20). Shang J et al. CircPAN3 contributes to drug resistance in acute myeloid leukemia through regulation of autophagy. Leuk Res 2019 Aug 2;85:106198 (PubMed: 31401408)

21). Zhao J et al. Inhibition of CCL19 benefits non‑alcoholic fatty liver disease by inhibiting TLR4/NF‑κB‑p65 signaling. Mol Med Rep 2018 Sep 14 (PubMed: 30221732)

22). Chen M;Xu J;Wang Y;Wang Z;Guo L;Li X;Huang L; et al. Arctium lappa L. polysaccharide can regulate lipid metabolism in type 2 diabetic rats through the SREBP-1/SCD-1 axis. Carbohydr Res 2020 Jun 7;494:108055. (PubMed: 32535406)

23). Feng Wang et al. CB2 receptor agonist JWH133 activates AMPK to inhibit growth of C6 glioma cells. OPEN LIFE SCI 2019

24). Liu N;Fu D;Yang J;Liu P;Song X;Wang X;Li R;Fu Z;Chen J;Gong X;Chen C;Yang L; et al. Asiatic acid attenuates hypertrophic and fibrotic differentiation of articular chondrocytes via AMPK/PI3K/AKT signaling pathway. Arthritis Res Ther 2020 May 12;22(1):112. (PubMed: 32398124)

25). Zhang DS et al. Role of Phosphorylated AMP-Activated Protein Kinase (AMPK) in Myocardial Insulin Resistance After Myocardial Ischemia-Reperfusion During Cardiopulmonary Bypass Surgery in Dogs. Med Sci Monit 2019 Jun 4;25:4149-4158 (PubMed: 31160548)

26). et al. Potential role of mTORC1 and the PI3K-Akt pathway in anti-acne properties of licorice flavonoids.

27). Ma MQ et al. A 6 hour therapeutic window, optimal for interventions targeting AMPK synergism and apoptosis antagonism, for cardioprotection against myocardial ischemic injury: an experimental study on rats. Am J Cardiovasc Dis 2015 Mar 20;5(1):63-71 (PubMed: 26064793)

28). et al. Hydrogen sulfide improves endothelial dysfunction in hypertension by activating peroxisome proliferatoractivated receptor delta/endothelial nitric oxide synthase signaling.

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Catalog Number :

AF3423-BP
(Blocking peptide available as AF3423-BP)

Price/Size :

$350/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Zebrafish
100%
Chicken
100%
Rabbit
100%
Xenopus
100%
Dog
100%
Bovine
100%
Sheep
100%
Pig
100%
Horse
0%
High similarity Medium similarity Low similarity No similarity
Q13131/P54646 as Substrate
Site PTM Type Enzyme
S6 Phosphorylation
T32 Phosphorylation
K40 Ubiquitination
K45 Ubiquitination
K52 Ubiquitination
K56 Ubiquitination
K71 Ubiquitination
K80 Acetylation
K80 Ubiquitination
K152 Ubiquitination
S172 Phosphorylation
S176 Phosphorylation
T183 Phosphorylation Q8TDC3-2 (BRSK1) , Q16584 (MAP3K11) , Q15831 (STK11) , Q13554 (CAMK2B) , Q8N5S9 (CAMKK1) , Q13131 (PRKAA1) , Q96RR4 (CAMKK2) , Q8IWQ3 (BRSK2)
S184 Phosphorylation
C185 S-Nitrosylation
S187 Phosphorylation
Y190 Phosphorylation
Y247 Phosphorylation
K266 Ubiquitination
K271 Ubiquitination
K280 Ubiquitination
K285 Ubiquitination
S293 Phosphorylation
Y294 Phosphorylation
T355 Phosphorylation
S356 Phosphorylation
S360 Phosphorylation Q13131 (PRKAA1) , O75385 (ULK1)
T368 Phosphorylation O75385 (ULK1)
T382 Phosphorylation
T388 Phosphorylation Q13131 (PRKAA1)
K396 Ubiquitination
S397 Phosphorylation O75385 (ULK1)
K406 Sumoylation
K406 Ubiquitination
K429 Ubiquitination
Y441 Phosphorylation
Y442 Phosphorylation
K448 Ubiquitination
Y463 Phosphorylation
S467 Phosphorylation
T482 Phosphorylation
K485 Ubiquitination
S486 Phosphorylation P49841 (GSK3B) , O75385 (ULK1) , Q13131 (PRKAA1) , P17612 (PRKACA)
T488 Phosphorylation O75385 (ULK1)
T490 Phosphorylation P49841 (GSK3B)
S494 Phosphorylation Q13131 (PRKAA1)
S496 Phosphorylation Q13131 (PRKAA1) , P31749 (AKT1)
S498 Phosphorylation
Y500 Phosphorylation
S506 Phosphorylation
S508 Phosphorylation
K515 Ubiquitination
S520 Phosphorylation
T522 Phosphorylation
S523 Phosphorylation
S524 Phosphorylation
T526 Phosphorylation
S527 Phosphorylation
S531 Phosphorylation
Site PTM Type Enzyme
K41 Ubiquitination
K45 Ubiquitination
K69 Acetylation
K69 Ubiquitination
K141 Ubiquitination
S161 Phosphorylation
S165 Phosphorylation
T172 Phosphorylation Q96RR4 (CAMKK2) , O43318 (MAP3K7) , Q15831 (STK11)
S173 Phosphorylation P17612 (PRKACA)
C174 S-Nitrosylation
S176 Phosphorylation
Y179 Phosphorylation
R227 Methylation
Y232 Phosphorylation
K260 Ubiquitination
K269 Ubiquitination
Y324 Phosphorylation
S377 Phosphorylation
K391 Ubiquitination
K401 Sumoylation
K401 Ubiquitination
Y436 Phosphorylation
S481 Phosphorylation
S483 Phosphorylation
T485 Phosphorylation Q15831 (STK11)
S489 Phosphorylation
S491 Phosphorylation P31749 (AKT1)
S500 Phosphorylation
S501 Phosphorylation
S509 Phosphorylation
S511 Phosphorylation
S515 Phosphorylation
T521 Phosphorylation
T524 Phosphorylation
S527 Phosphorylation
S534 Phosphorylation
Q13131/P54646 as PTM Enzyme
Substrate Site Source
O00418 (EEF2K) S78 Uniprot
O00418 (EEF2K) S366 Uniprot
O00418 (EEF2K) S398 Uniprot
O00429 (DNM1L) S637 Uniprot
O00763 (ACACB) S222 Uniprot
O15350 (TP73) S426 Uniprot
O15360 (FANCA) S347 Uniprot
O43524 (FOXO3) T179 Uniprot
O43524 (FOXO3) S399 Uniprot
O43524 (FOXO3) S413 Uniprot
O43524 (FOXO3) S555 Uniprot
O43524 (FOXO3) S588 Uniprot
O43524 (FOXO3) S626 Uniprot
O60825 (PFKFB2) S466 Uniprot
O75385 (ULK1) S317 Uniprot
O75385 (ULK1) S638 Uniprot
O95278 (EPM2A) S25 Uniprot
O95863 (SNAI1) S11 Uniprot
O95863 (SNAI1) S92 Uniprot
P00533 (EGFR) T892 Uniprot
P04049 (RAF1) S259 Uniprot
P04049 (RAF1) S621 Uniprot
P04406 (GAPDH) S122 Uniprot
P04626 (ERBB2) T900 Uniprot
P04637 (TP53) S15 Uniprot
P04637 (TP53) T18 Uniprot
P04637 (TP53) S20 Uniprot
P06400 (RB1) S811 Uniprot
P10636-8 (MAPT) S214 Uniprot
P10636-8 (MAPT) T231 Uniprot
P10636 (MAPT) S255 Uniprot
P10636-8 (MAPT) S262 Uniprot
P10636 (MAPT) S355 Uniprot
P10636-8 (MAPT) S356 Uniprot
P10636 (MAPT) S396 Uniprot
P10636-8 (MAPT) S422 Uniprot
P13569 (CFTR) S737 Uniprot
P13569 (CFTR) S768 Uniprot
P13569 (CFTR) S813 Uniprot
P14859 (POU2F1) S335 Uniprot
P14859 (POU2F1) S385 Uniprot
P15056 (BRAF) S729 Uniprot
P15531 (NME1) S122 Uniprot
P15531 (NME1) S144 Uniprot
P17600 (SYN1) S9 Uniprot
P19429 (TNNI3) S23 Uniprot
P19429 (TNNI3) S24 Uniprot
P19429 (TNNI3) S150 Uniprot
P29474 (NOS3) T495 Uniprot
P29474 (NOS3) S633 Uniprot
P29474 (NOS3) S1177 Uniprot
P36956-3 (SREBF1) S372 Uniprot
P41235 (HNF4A) S303 Uniprot
P41235 (HNF4A) S313 Uniprot
P42345 (MTOR) T2446 Uniprot
P43405 (SYK) S178 Uniprot
P46527 (CDKN1B) T198 Uniprot
P46937 (YAP1) S61 Uniprot
P46937 (YAP1) S94 Uniprot
P49116 (NR2C2) S351 Uniprot
P49674 (CSNK1E) S389 Uniprot
P50552 (VASP) T278 Uniprot
P52292 (KPNA2) S105 Uniprot
P54840 (GYS2) S8 Uniprot
P55011 (SLC12A2) S77 Uniprot
P63244 (RACK1) T50 Uniprot
Q04759 (PRKCQ) T538 Uniprot
Q05469 (LIPE) S855 Uniprot
Q06210 (GFPT1) S242 Uniprot
Q06210-2 (GFPT1) S243 Uniprot
Q06210 (GFPT1) S261 Uniprot
Q07866 (KLC1) S521 Uniprot
Q09472 (EP300) S89 Uniprot
Q12778 (FOXO1) T649 Uniprot
Q13002-1 (GRIK2) S715 Uniprot
Q13085-2 (ACACA) S22 Uniprot
Q13085 (ACACA) S80 Uniprot
Q13085-4 (ACACA) S117 Uniprot
Q13085-1 (ACACA) S1201 Uniprot
Q13085-4 (ACACA) S1238 Uniprot
Q13131 (PRKAA1) T183 Uniprot
Q13131 (PRKAA1) S360 Uniprot
Q13131 (PRKAA1) T388 Uniprot
Q13131 (PRKAA1) S486 Uniprot
Q13131 (PRKAA1) S494 Uniprot
Q13131 (PRKAA1) S496 Uniprot
Q13362 (PPP2R5C) S298 Uniprot
Q13362 (PPP2R5C) S336 Uniprot
Q13363 (CTBP1) S158 Uniprot
Q13621 (SLC12A1) S122 Uniprot
Q13621 (SLC12A1) S130 Uniprot
Q15036 (SNX17) S437 Uniprot
Q16526 (CRY1) S71 Uniprot
Q16875 (PFKFB3) S461 Uniprot
Q53ET0 (CRTC2) S170 Uniprot
Q6N021 (TET2) S99 Uniprot
Q7Z3C6 (ATG9A) S761 Uniprot
Q7Z628 (NET1) S100 Uniprot
Q86TI0 (TBC1D1) S237 Uniprot
Q86TI0 (TBC1D1) T596 Uniprot
Q8IXJ6 (SIRT2) T101 Uniprot
Q8N122 (RPTOR) S722 Uniprot
Q8N122 (RPTOR) S792 Uniprot
Q8WUI4 (HDAC7) S155 Uniprot
Q8WUI4 (HDAC7) S358 Uniprot
Q92538 (GBF1) T1337 Uniprot
Q9BU19 (ZNF692) S470 Uniprot
Q9BZL4 (PPP1R12C) S452 Uniprot
Q9GZY8 (MFF) S155 Uniprot
Q9GZY8 (MFF) S172 Uniprot
Q9H0B6 (KLC2) S539 Uniprot
Q9H0B6 (KLC2) S545 Uniprot
Q9H0B6 (KLC2) S581 Uniprot
Q9H0B6 (KLC2) S582 Uniprot
Q9NYV6 (RRN3) S635 Uniprot
Q9P2M7 (CGN) S131 Uniprot
Q9UBK2 (PPARGC1A) T178 Uniprot
Q9UQK1 (PPP1R3C) S33 Uniprot
Q9UQK1 (PPP1R3C) S293 Uniprot
Q9UQL6 (HDAC5) S259 Uniprot
Q9UQL6 (HDAC5) S498 Uniprot
Q9Y478 (PRKAB1) S24 Uniprot
Q9Y478 (PRKAB1) T80 Uniprot
Q9Y478 (PRKAB1) S108 Uniprot
Q9Y478 (PRKAB1) T148 Uniprot
Q9Y478 (PRKAB1) T158 Uniprot
Q9Y478 (PRKAB1) S174 Uniprot
Q9Y478 (PRKAB1) S177 Uniprot
Substrate Site Source
F1D8S2 (NR2A1) S313 Uniprot
O00418 (EEF2K) S78 Uniprot
O00418 (EEF2K) S366 Uniprot
O00418 (EEF2K) S398 Uniprot
O00763-1 (ACACB) S222 Uniprot
O14920 (IKBKB) S177 Uniprot
O14920 (IKBKB) S181 Uniprot
O15151 (MDM4) S342 Uniprot
O43524 (FOXO3) T179 Uniprot
O43524 (FOXO3) S399 Uniprot
O43524 (FOXO3) S413 Uniprot
O43524 (FOXO3) S555 Uniprot
O43524 (FOXO3) S588 Uniprot
O43524 (FOXO3) S626 Uniprot
O60825 (PFKFB2) S466 Uniprot
O75385 (ULK1) S317 Uniprot
O75385 (ULK1) S556 Uniprot
O75385 (ULK1) S638 Uniprot
P05549 (TFAP2A) S219 Uniprot
P06241 (FYN) T12 Uniprot
P08151 (GLI1) S102 Uniprot
P08151 (GLI1) S408 Uniprot
P08151 (GLI1) T1074 Uniprot
P09874 (PARP1) S177 Uniprot
P28562 (DUSP1) S334 Uniprot
P29474 (NOS3) S633 Uniprot
P29474 (NOS3) S1177 Uniprot
P30260 (CDC27) S379 Uniprot
P35222 (CTNNB1) S552 Uniprot
P36956 (SREBF1) S396 Uniprot
P41235-3 (HNF4A) S313 Uniprot
P49815 (TSC2) S1387 Uniprot
P50552 (VASP) T278 Uniprot
P50552 (VASP) S322 Uniprot
P55011 (SLC12A2) S77 Uniprot
P55011 (SLC12A2) S242 Uniprot
Q13085-2 (ACACA) S22 Uniprot
Q13085 (ACACA) S78 Uniprot
Q13085 (ACACA) S80 Uniprot
Q13085-4 (ACACA) S117 Uniprot
Q13177 (PAK2) S20 Uniprot
Q13393 (PLD1) S505 Uniprot
Q15121 (PEA15) S116 Uniprot
Q53ET0 (CRTC2) S171 Uniprot
Q86TI0 (TBC1D1) S237 Uniprot
Q8IY63 (AMOTL1) S793 Uniprot
Q8N122 (RPTOR) S863 Uniprot
Q8NFG4 (FLCN) S62 Uniprot
Q92819 (HAS2) T110 Uniprot
Q96EB6 (SIRT1) T344 Uniprot
Q9BZL4 (PPP1R12C) S452 Uniprot
Q9UQB8 (BAIAP2) S366 Uniprot
Q9UQL6 (HDAC5) S259 Uniprot
Q9UQL6 (HDAC5) S498 Uniprot
Q9Y2I7 (PIKFYVE) S307 Uniprot
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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