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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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Agonist-induced Serine403/Serine404 phosphorylation of the Neurotensin Receptor 1
pS403/pS404-NTS1 (phospho-Neurotensin Receptor...
Serine403/Serine404 (S403/S404) is major phosphorylation site of the Neurotensin Receptor 1 (NTS1). The pS403/pS404-NTS1 antibody detects phosphorylation in response to agonists. S403/S404 phosphorylation is likely to be involved in...
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NEW
Agonist-induced Threonine382/Threonine386 phosphorylation of the Dopamine Receptor 5
pT382/pT386-D5 (phospho-Dopamine Receptor 5...
Threonine382/Threonine386 (T382/T386) is a major phosphorylation site of the D5 receptor. The pT382/pT386-D5 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T382/T386...
$ 375.00 *
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Agonist-induced Serine377/Ser378 phosphorylation of the Oxytocin Receptor.
pS377/pS378-OT (phospho-Oxytocin Receptor...
Serine377/Serine378 (S377/S378) is major phosphorylation site of the Oxytocin Receptor (OT). The p3S77/pS378-OT antibody detects phosphorylation in response to agonists. S377/S378 phosphorylation is likely to be involved in efficient...
$ 375.00 *
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Agonist-induced Serine366/Serine368 phosphorylation of the Oxytocin Receptor
pS366/pS368-OT (phospho-Oxytocin Receptor...
Serine366/Serine368 (S366/S368) is major phosphorylation site of the Oxytocin Receptor (OT). The pS366/pS368-OT antibody detects phosphorylation in response to agonists. S366/S368 phosphorylation is likely to be involved in efficient...
$ 375.00 *
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Agonist-induced Threonine310/Serine311 phosphorylation of the M2 Muscarinic Acetycholine Receptor
pT310/pS311-M2 (phospho-M2 Muscarinic...
Threonine310/Serine311 is a major phosphorylation site of the M2 Muscarinic Acetylcholine Receptor. The pT310/pS311-M2 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T310/S311...
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Agonist-induced Serine315/Serine320 phosphorylation of the M2 Muscarinic Acetycholine Receptor
pS315/pS320-M2 (phospho-M2 Muscarinic...
Serine315/Serine320 is a major phosphorylation site of the M2 Muscarinic Acetylcholine Receptor. The pS315/pS320-M2 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S315/S320...
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Agonist-induced Serine286/Threonine287/Serine288 phosphorylation of the M2 Muscarinic Acetycholine Receptor
pS286/pT287/pS288-M2 (phospho-M2 Muscarinic...
Serine286/Threonine287/Serine288 is a major phosphorylation site of the M2 Muscarinic Acetylcholine Receptor. The pS286/pT287/pS288-M2 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation....
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Validation of the δ-Opioid Receptor in transfected HEK293 cells
DOP (non-phospho) ∂-Opioid Receptor Antibody
The non-phospho-∂-opioid receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human DOP. It can be used to detect total DOP receptors in Western blots independent of phosphorylation.
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Validation of the Complement C5a Receptor 2 in transfected HEK293 cells.
C5a2 (non-phospho), Complement C5a Receptor 2...
The non-phospho-C5a2 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human C5a2. It can be used to detect total C5a2 receptors in Western blots independent of phosphorylation. The non-phospho-C5a2...
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Validation of the Atypical Chemokine Receptor 2 in transfected HEK293 cells.
ACKR2 (non-phospho), Atypical Chemokine...
The non-phospho-ACKR2 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human ACKR2. It can be used to detect total ACKR2 receptors in Western blots independent of phosphorylation. The...
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Validation of the β2-Adrenoceptor in transfected HEK293 cells
β2 (non-phospho), β2-Adrenoceptor Antibody
The non-phospho- β2 antibody is directed against the third intracellular loop of human β2 . It can be used to detect total β2 receptors in Western blots independent of phosphorylation. The non-phospho- β2 antibody can also be used to...
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Agonist-induced Serine369/Serine374 phosphorylation of the Neuropeptide Y receptor 2
pS369/pS374-NPY2 (phospho-Neuropeptide Y...
Serine369/Serine374 (S369/S374) is a major phosphorylation site of the Neuropeptide Y Receptor 2. The pS369/pS374-NPY2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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Agonist-induced Serine354/Threonine356 phosphorylation of the Neuropeptide Y receptor 2
pS354/pT356-NPY2 (phospho-Neuropeptide Y...
Serine354Threonine356 (S354/T356) is a major phosphorylation site of the Neuropeptide Y Receptor 2. The pS354/pT356-NPY2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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NEW
Agonist-induced Serine351/Serine354 phosphorylation of the Neuropeptide Y
pS351/pS354-NPY2 (phospho-Neuropeptide Y...
Serine351/Serine354 (S351/S354) is a major phosphorylation site of the Neuropeptide Y Receptor 2. The pS351/pS354-NPY2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
$ 375.00 *
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NEW
Agonist-induced Threonine355 phosphorylation of the Chemokine Receptor 10
pT355-CCR10 (phospho-Chemokine Receptor 10...
Threonine355 (T355) is a major phosphorylation site of the CCR10 receptor. The pT355-CCR10 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T355 phosphorylation is a key regulator...
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Agonist-induced Threonine271 phosphorylation of the M2 Muscarinic Acetycholine Receptor
M2 Sample Pack (phospho- and non-phospho-M2...
M2 Sample Pack consisting of all six available phospho- and one non-phospho-M2 Receptor Antibodies 7 x 20 µL trial size each. Specifically, this sample pack contains the following antibodies pT271-M2 (7TM0014A), pS282/pS283-M2...
$ 300.00 *
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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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Premium Phosphosite-Specific 7TM Antibodies

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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