Product: Phospho-AMPK alpha 1 (Ser486) Antibody
Catalog: AF3422
Description: Rabbit polyclonal antibody to Phospho-AMPK alpha 1 (Ser486)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Zebrafish, Bovine, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 62kDa, 70 kDa; 64kD(Calculated).
Uniprot: Q13131
RRID: AB_2834864

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Zebrafish(82%), Bovine(100%), Sheep(100%), Rabbit(91%), Dog(100%), Chicken(100%), Xenopus(80%)
Clonality:
Polyclonal
Specificity:
Phospho-AMPK alpha 1 (Ser486) Antibody detects endogenous levels of AMPK alpha 1 only when phosphorylated at Ser486(Isoform 1).
RRID:
AB_2834864
Cite Format: Affinity Biosciences Cat# AF3422, RRID:AB_2834864.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
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.
Alias:

Fold/Unfold

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;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Description:
AMPKA1 a protein kinase of the CAMKL family that plays a central role in regulating cellular and organismal energy balance in response to the balance between AMP/ATP, and intracellular Ca(2+) levels.
Sequence:
MRRLSSWRKMATAEKQKHDGRVKIGHYILGDTLGVGTFGKVKVGKHELTGHKVAVKILNRQKIRSLDVVGKIRREIQNLKLFRHPHIIKLYQVISTPSDIFMVMEYVSGGELFDYICKNGRLDEKESRRLFQQILSGVDYCHRHMVVHRDLKPENVLLDAHMNAKIADFGLSNMMSDGEFLRTSCGSPNYAAPEVISGRLYAGPEVDIWSSGVILYALLCGTLPFDDDHVPTLFKKICDGIFYTPQYLNPSVISLLKHMLQVDPMKRATIKDIREHEWFKQDLPKYLFPEDPSYSSTMIDDEALKEVCEKFECSEEEVLSCLYNRNHQDPLAVAYHLIIDNRRIMNEAKDFYLATSPPDSFLDDHHLTRPHPERVPFLVAETPRARHTLDELNPQKSKHQGVRKAKWHLGIRSQSRPNDIMAEVCRAIKQLDYEWKVVNPYYLRVRRKNPVTSTYSKMSLQLYQVDSRTYLLDFRSIDDEITEAKSGTATPQRSGSVSNYRSCQRSDSDAEAQGKSSEVSLTSSVTSLDSSPVDLTPRPGSHTIEFFEMCANLIKILAQ

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Bovine
100
Sheep
100
Dog
100
Chicken
100
Rabbit
91
Zebrafish
82
Xenopus
80
Pig
0
Horse
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q13131 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

PTMs - Q13131 As 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

Research Backgrounds

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. 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.

PTMs:

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 assertionSubcellular location
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.

Family&Domains:

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

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (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 > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Apelin signaling pathway.   (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

· Human Diseases > Cardiovascular diseases > Hypertrophic cardiomyopathy (HCM).

· Organismal Systems > Aging > Longevity regulating pathway.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway - multiple species.   (View pathway)

· Organismal Systems > Environmental adaptation > Circadian rhythm.   (View pathway)

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

· Organismal Systems > Endocrine system > Glucagon signaling pathway.

References

1). [EXPRESS] mTOR activation due to APPL1 deficiency exacerbates hyperalgesia via Rab5/Akt and AMPK signaling pathway in STZ-induced diabetic rats. Molecular Pain, 2019 (PubMed: 31530215) [IF=3.3]

Application: WB    Species: rat    Sample:

Figure 7.| The involvement of AMPK and Akt in the negative regulation of mTOR by APPL1 in STZ-induced diabetic rats. The variations of p-AMPK and p-Akt in normal control rats with vacant lentiviral vector control, normal control rats with APPL1 knockdown, PDN rats, and PDN rats with APPL1 knockdown, and PDN rats with APPL1 overexpression. The abbreviations for the groups of negative control, normal control + shRNA encoding APPL1, painful diabetic neuropathy (PDN), PDN + shRNA encoding APPLL (shRNA) and PDN + APPL1 genetic overexpression are shown as CON, CON + shRNA, PDN, PDN + shRNA, and PDN + OXP. (n = 4, *P < 0.05 vs. NC group, # P < 0.05 vs. PDN group). Data are expressed as the means ± SEM.

2). Liposomal Silybin Improves Glucose and Lipid Metabolisms in Type 2 Diabetes Mellitus Complicated with Non-Alcoholic Fatty Liver Disease via AMPK/TGF-β1/Smad Signaling. The Tohoku journal of experimental medicine, 2023 (PubMed: 37344419) [IF=2.2]

3). Metformin Combined with Alendronate Ameliorates Osteoarthritis by Attenuating RANKL-induced Bone Resorption and Protecting Cartilage Against Degradation. Research Square, 2022

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