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Phosphorylation of intracellular serine and threonine residues is the most important post translational modification of G protein-coupled receptors (GPCRs) also called heptahelical or seven transmembrane receptors (7TMR). After agonist exposure, these receptors acquire an active conformation, which is recognized by a family of highly specialized GPCR kinases (GRKs). Agonist-driven phosphorylation by GRKs regulates acute receptor desensitization, arrestin recruitment, internalization, post-activation signaling, long-term tolerance and drug addiction. Phosphosite-specific 7TM antibodies are designed to specifically detect agonist-activated GPCRs. In fact, recent work shows that ligand profiling using phosphosite-specific 7TM antibodies provides valuble information on ligand bias beyond that obtained with conventional ß-arrestin recruitment assays. Phosphosite-specific 7TM antibodies are novel tools for GPCR research that can be used to:

  • profile agonist properties of novel GPCR ligands
  • decipher the phosphorylation barcode of GPCRs
  • determine the spatial and temporal dynamics of receptor phosphorylation
  • identify relevant kinases and phosphatases for GPCR phosphorylation and dephosphoryation

Lifecycle3

Schematic representation of the G protein-coupled receptor phosphorylation / dephosphorylation cycle. GRK, G protein-coupled receptor kinase; PKC, protein kinase C; cPP1, catalytic subunit of protein phosphatase 1; R*, activated GPCR; CCP, clathrin-coated pit. 

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Validation of the Calcium-sensing Receptor in transfected HEK293 cells
CaS (non-phospho), Calcium-sensing Receptor...
The non-phospho-CaS Calcium-sensing Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human CaS. It can be used to detect total CaS receptors in Western blots independent of phosphorylation. The...
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Immunohistochemical identification of Calcium-sensing Receptor in pancreas
CaS (IHC-grade), Calcium-sensing Receptor Antibody
The non-phospho-CaS Calcium-sensing Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human CaS. It can be used to detect total CaS receptors in Western blots independent of phosphorylation. The...
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pT311/pS314-M4 (phospho-M4 Muscarinic Acetylcholine Receptor Antibody)
pT311/pS314-M4 (phospho-M4 Muscarinic...
Threonine311/Serine314 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pT311/pS314-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T311/S314...
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pS296/pS297-M4 (phospho-M4 Muscarinic Acetylcholine Receptor Antibody)
pS296/pS297-M4 (phospho-M4 Muscarinic...
Serine296/Serine297 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pS296/pS297-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S296/S297...
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Agonist-induced Threonine315/Threonine316 phosphorylation of the M4 Muscarinic Acetylcholine Receptor
pT315/pT316-M4 (phospho-M4 Muscarinic...
Threonine315/Threonine316 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pT315/pT316-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T315/T316...
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Agonist-induced Serine298/Serine300 phosphorylation of the M4 Muscarinic Acetylcholine Receptor
pS298/pS300-M4 (phospho-M4 Muscarinic...
Serine298/Serine300 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pS298/pS300-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S298/S300...
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Immunohistochemical identification of Serine298/Serine300 phosphorylation of M4 Receptor in striatum
pS298/pS300-M4 (IHC-grade phospho-M4 Muscarinic...
Serine298/Serine300 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pS298/pS300-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S298/S300...
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Agonist-induced Serine473/Serine475 phosphorylation of the β1-Adrenoceptor
pS473/pS475-β1 (phospho-β1-Adrenoceptor Antibody)
Serine473/Serine475 (S473/S475) is a major phosphorylation site of the β1 adrenoceptor. The pS473/pS475-β1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S473/S475...
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Isolation and enrichment of the Smoothened Receptor from tissue lysates
SMO (GP-non-phospho) Smoothened Receptor...
The non-phospho SMO receptor antibody is directed against the distal end of the carboxyl-terminal tail human SMO receptor. It can be used to detect total SMO receptors in Western blots independent of phosphorylation. It can also be used...
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Immunohistochemical identification of Smoothened Receptor in heart
SMO (GP-IHC-grade), Smoothened Receptor...
The non-phospho SMO receptor antibody is directed against the distal end of the carboxyl-terminal tail human SMO receptor. It can be used to detect total SMO receptors in Western blots independent of phosphorylation. It can also be used...
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Immunohistochemical identification of Smoothened Receptor in cilia
SMO (IHC-grade), Smoothened Receptor Antibody
The non-phospho SMO receptor antibody is directed against the distal end of the carboxyl-terminal tail human SMO receptor. It can be used to detect total SMO receptors in Western blots independent of phosphorylation. It can also be used...
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Validation of the Smoothened Receptor in transfected HEK293 cells
SMO (non-phospho) Smoothened Receptor Antibody
The non-phospho SMO receptor antibody is directed against the distal end of the carboxyl-terminal tail human SMO receptor. It can be used to detect total SMO receptors in Western blots independent of phosphorylation. It can also be used...
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Validation of the Melanin-Concentrating Hormone Receptor 2 in transfected HEK293 cells
MCH2 (non-phospho) Melanin-Concentrating...
The non-phospho MCH2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human MCH2 receptor. It can be used to detect total MCH2 receptors in Western blots independent of phosphorylation. It can also be...
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Validation of the A2A Adenosine Receptor in transfected HEK293 cells
A2A (GP-non-phospho) Adenosine 2A Receptor...
The non-phospho-A2A receptor antibody is directed against the distal end of the carboxyl-terminal tail of human A2A. It can be used to detect total A2A receptors in Western blots independent of phosphorylation. The A2A antibody can also...
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Validation of the Hydroxycarboxylic Acid Receptor 2 in transfected HEK293 cells
HCA2 (non-phospho) Hydroxycarboxylic Acid...
The non-phospho HCA2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human HCA2 receptor. It can be used to detect total HCA2 receptors in Western blots independent of phosphorylation. It can also be...
375.00 € *
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Validation of the Hydroxycarboxylic Acid Receptor 2 in transfected HEK293 cells
HCA2 (GP-non-phospho) Hydroxycarboxylic Acid...
The non-phospho HCA2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human HCA2 receptor. It can be used to detect total HCA2 receptors in Western blots independent of phosphorylation. It can also be...
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For further reading refer to:

Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636. doi:10.1016/j.tips.2017.04.002. Epub 2017 May 4. Review. PubMed PMID: 28478994.

Miess E, Gondin AB, Yousuf A, Steinborn R, Mösslein N, Yang Y, Göldner M, Ruland JG, Bünemann M, Krasel C, Christie MJ, Halls ML, Schulz S, Canals M. Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid μ-opioid receptor desensitization. Sci Signal. 2018 Jul 17;11(539). pii: eaas9609. doi: 10.1126/scisignal.aas9609. PubMed PMID: 30018083.

Kliewer A, Schmiedel F, Sianati S, Bailey A, Bateman JT, Levitt ES, Williams JT, Christie MJ, Schulz S. Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia and diminish tolerance but worsen opioid side effects. Nat Commun. 2019 Jan 21;10(1):367. doi: 10.1038/s41467-018-08162-1. PubMed PMID: 30664663; PubMed Central PMCID: PMC6341117.

Mann A, Moulédous L, Froment C, O'Neill PR, Dasgupta P, Günther T, Brunori G, Kieffer BL, Toll L, Bruchas MR, Zaveri NT, Schulz S. Agonist-selective NOP receptor phosphorylation correlates in vitro and in vivo and reveals differential post-activation signaling by chemically diverse agonists. Sci Signal. 2019 Mar 26;12(574). pii: eaau8072. doi: 10.1126/scisignal.aau8072. PubMed PMID: 30914485; PubMed Central PMCID: PMC6934085.

Saaber F, Schütz D, Miess E, Abe P, Desikan S, Ashok Kumar P, Balk S, Huang K, Beaulieu JM, Schulz S, Stumm R. ACKR3 Regulation of Neuronal Migration Requires ACKR3 Phosphorylation, but Not β-Arrestin. Cell Rep. 2019 Feb 5;26(6):1473-1488.e9. doi: 10.1016/j.celrep.2019.01.049. PubMed PMID: 30726732.

Glück L, Loktev A, Moulédous L, Mollereau C, Law PY, Schulz S. Loss of morphine reward and dependence in mice lacking G protein-coupled receptor kinase 5. Biol Psychiatry. 2014 Nov 15;76(10):767-74. doi: 10.1016/j.biopsych.2014.01.021. Epub 2014 Feb 3. PubMed PMID: 24629717; PubMed Central PMCID: PMC4119866.

For further reading refer to: Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636.... read more »
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For further reading refer to:

Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636. doi:10.1016/j.tips.2017.04.002. Epub 2017 May 4. Review. PubMed PMID: 28478994.

Miess E, Gondin AB, Yousuf A, Steinborn R, Mösslein N, Yang Y, Göldner M, Ruland JG, Bünemann M, Krasel C, Christie MJ, Halls ML, Schulz S, Canals M. Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid μ-opioid receptor desensitization. Sci Signal. 2018 Jul 17;11(539). pii: eaas9609. doi: 10.1126/scisignal.aas9609. PubMed PMID: 30018083.

Kliewer A, Schmiedel F, Sianati S, Bailey A, Bateman JT, Levitt ES, Williams JT, Christie MJ, Schulz S. Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia and diminish tolerance but worsen opioid side effects. Nat Commun. 2019 Jan 21;10(1):367. doi: 10.1038/s41467-018-08162-1. PubMed PMID: 30664663; PubMed Central PMCID: PMC6341117.

Mann A, Moulédous L, Froment C, O'Neill PR, Dasgupta P, Günther T, Brunori G, Kieffer BL, Toll L, Bruchas MR, Zaveri NT, Schulz S. Agonist-selective NOP receptor phosphorylation correlates in vitro and in vivo and reveals differential post-activation signaling by chemically diverse agonists. Sci Signal. 2019 Mar 26;12(574). pii: eaau8072. doi: 10.1126/scisignal.aau8072. PubMed PMID: 30914485; PubMed Central PMCID: PMC6934085.

Saaber F, Schütz D, Miess E, Abe P, Desikan S, Ashok Kumar P, Balk S, Huang K, Beaulieu JM, Schulz S, Stumm R. ACKR3 Regulation of Neuronal Migration Requires ACKR3 Phosphorylation, but Not β-Arrestin. Cell Rep. 2019 Feb 5;26(6):1473-1488.e9. doi: 10.1016/j.celrep.2019.01.049. PubMed PMID: 30726732.

Glück L, Loktev A, Moulédous L, Mollereau C, Law PY, Schulz S. Loss of morphine reward and dependence in mice lacking G protein-coupled receptor kinase 5. Biol Psychiatry. 2014 Nov 15;76(10):767-74. doi: 10.1016/j.biopsych.2014.01.021. Epub 2014 Feb 3. PubMed PMID: 24629717; PubMed Central PMCID: PMC4119866.

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