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PerkinElmer damgo
Association kinetics of δOR radioligand agonist binding. SK-N-SH whole cells endogenously expressing μOR and δOR (2:1 ratio) were incubated with 6 nM [ 3 <t>H]deltorphin</t> II in the absence or presence of 10 nM <t>DAMGO</t> or fentanyl for different
Damgo, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Price from $9.99 to $1999.99
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Images

1) Product Images from "G Protein-Coupled Receptor Heteromerization: A Role in Allosteric Modulation of Ligand Binding S⃞"

Article Title: G Protein-Coupled Receptor Heteromerization: A Role in Allosteric Modulation of Ligand Binding S⃞

Journal: Molecular Pharmacology

doi: 10.1124/mol.110.070847

Association kinetics of δOR radioligand agonist binding. SK-N-SH whole cells endogenously expressing μOR and δOR (2:1 ratio) were incubated with 6 nM [ 3 H]deltorphin II in the absence or presence of 10 nM DAMGO or fentanyl for different
Figure Legend Snippet: Association kinetics of δOR radioligand agonist binding. SK-N-SH whole cells endogenously expressing μOR and δOR (2:1 ratio) were incubated with 6 nM [ 3 H]deltorphin II in the absence or presence of 10 nM DAMGO or fentanyl for different

Techniques Used: Binding Assay, Expressing, Incubation

Dissociation kinetics of [ 3 H]deltorphin II (A) or [ 3 H]DAMGO (B) binding. A, SK-N-SH whole cells endogenously expressing μOR and δOR were incubated with 6 nM [ 3 H]deltorphin II for 1 h at 37°C. The supernatant was removed, the plates
Figure Legend Snippet: Dissociation kinetics of [ 3 H]deltorphin II (A) or [ 3 H]DAMGO (B) binding. A, SK-N-SH whole cells endogenously expressing μOR and δOR were incubated with 6 nM [ 3 H]deltorphin II for 1 h at 37°C. The supernatant was removed, the plates

Techniques Used: Binding Assay, Expressing, Incubation

2) Product Images from "Dopamine D4 Receptor Counteracts Morphine-Induced Changes in ? Opioid Receptor Signaling in the Striosomes of the Rat Caudate Putamen"

Article Title: Dopamine D4 Receptor Counteracts Morphine-Induced Changes in ? Opioid Receptor Signaling in the Striosomes of the Rat Caudate Putamen

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms15011481

D 4 R in vitro activation prevents morphine-induced changes on MOR-dependent [ 35 S]GTPγS binding. ( A – D3 ) Representative autoradiograms of [ 35 S]GTPγS binding in coronal sections of rat brain at the CPu level. Rats were continuously treated with vehicle ( A – A3 ); morphine (20 mg/kg/day) ( B – B3 ); PD168,077 (1 mg/kg/day) ( C – C3 ) and morphine + PD168,077 ( D – D3 ); Basal levels of [ 35 S]GTPγS binding was determined in control sections from the four treatment groups ( A , B , C , D ) and in vitro receptor stimulation was performed with DAMGO (3 μM) ( A1 , B1 , C1 , D1 ); PD168,077 (90 nM) ( A2 , B2 , C2 , D2 ) or DAMGO + PD168,077 ( A3 , B3 , C3 , D3 ). Scale bar is 2 mm; ( E ) Effect of continuous drug treatments on [ 35 S]GTPγS binding in the rat CPu after in vitro agonist stimulation of MOR and/or D 4 R. Data represent mean ± SEM ( n = 6) and are expressed as percentage of basal [ 35 S]GTPγS binding value in vehicle-treated animals (red line). Blue line represents DAMGO-dependent [ 35 S]GTPγS binding in vehicle-treated animals. Differences between groups were set by two-way ANOVA followed by post hoc Bonferroni t test. * p
Figure Legend Snippet: D 4 R in vitro activation prevents morphine-induced changes on MOR-dependent [ 35 S]GTPγS binding. ( A – D3 ) Representative autoradiograms of [ 35 S]GTPγS binding in coronal sections of rat brain at the CPu level. Rats were continuously treated with vehicle ( A – A3 ); morphine (20 mg/kg/day) ( B – B3 ); PD168,077 (1 mg/kg/day) ( C – C3 ) and morphine + PD168,077 ( D – D3 ); Basal levels of [ 35 S]GTPγS binding was determined in control sections from the four treatment groups ( A , B , C , D ) and in vitro receptor stimulation was performed with DAMGO (3 μM) ( A1 , B1 , C1 , D1 ); PD168,077 (90 nM) ( A2 , B2 , C2 , D2 ) or DAMGO + PD168,077 ( A3 , B3 , C3 , D3 ). Scale bar is 2 mm; ( E ) Effect of continuous drug treatments on [ 35 S]GTPγS binding in the rat CPu after in vitro agonist stimulation of MOR and/or D 4 R. Data represent mean ± SEM ( n = 6) and are expressed as percentage of basal [ 35 S]GTPγS binding value in vehicle-treated animals (red line). Blue line represents DAMGO-dependent [ 35 S]GTPγS binding in vehicle-treated animals. Differences between groups were set by two-way ANOVA followed by post hoc Bonferroni t test. * p

Techniques Used: In Vitro, Activation Assay, Binding Assay

3) Product Images from "Mu-Opioid Receptor Coupling to G?o Plays an Important Role in Opioid Antinociception"

Article Title: Mu-Opioid Receptor Coupling to G?o Plays an Important Role in Opioid Antinociception

Journal: Neuropsychopharmacology

doi: 10.1038/npp.2011.91

DAMGO- and morphine-stimulated G protein activity in spinal cord homogenates from Gα o transgenic mice. [ 35 S]GTPγS (0.1 nM) incorporation stimulated by 10 μM DAMGO or morphine was evaluated in membrane homogenates from spinal cord of wild-type and Gα o +/− mice. Nonspecific binding was evaluated in the presence of unlabeled GTPγS (10 μM). Data are plotted as agonist-stimulated [ 35 S]GTPγS binding, defined as the increase in [ 35 S]GTPγS binding in the presence of agonist over that of basal (measured in the absence of agonist), and represent the mean±SEM ( n =3 performed in quadruplicate). Asterisks indicate a statistical difference vs wild type by Student's paired t -test ( * p
Figure Legend Snippet: DAMGO- and morphine-stimulated G protein activity in spinal cord homogenates from Gα o transgenic mice. [ 35 S]GTPγS (0.1 nM) incorporation stimulated by 10 μM DAMGO or morphine was evaluated in membrane homogenates from spinal cord of wild-type and Gα o +/− mice. Nonspecific binding was evaluated in the presence of unlabeled GTPγS (10 μM). Data are plotted as agonist-stimulated [ 35 S]GTPγS binding, defined as the increase in [ 35 S]GTPγS binding in the presence of agonist over that of basal (measured in the absence of agonist), and represent the mean±SEM ( n =3 performed in quadruplicate). Asterisks indicate a statistical difference vs wild type by Student's paired t -test ( * p

Techniques Used: Activity Assay, Transgenic Assay, Mouse Assay, Binding Assay

4) Product Images from "A mu–delta opioid receptor brain atlas reveals neuronal co-occurrence in subcortical networks"

Article Title: A mu–delta opioid receptor brain atlas reveals neuronal co-occurrence in subcortical networks

Journal: Brain Structure & Function

doi: 10.1007/s00429-014-0717-9

Expression of functional receptors in MOR-mcherry knock-in mice. a Targeting strategy: Oprm1 exons, mcherry cDNA, and the FRT ( triangle ) flanked neomycin cassette are, respectively, displayed as exon number , mcherry , and neo . Homologous recombination (HR) was followed by FLP recombinase treatment (FLP) in ES cells. Positions of the oligonucleotides (BAZ 43, BAZ 44) used for genotyping are indicated. b Western blot: detection of MOR-mcherry fusion by immunoblotting with antibodies directed against mcherry on membranes from striatum and periaqueductal gray (PAG) from wild-type ( Oprm1 +/+ ), heterozygote ( Oprm1 +/mch ) and homozygote ( Oprm1 mch/mch ) mice (MOR-mcherry fusion indicated by arrow ). Cos cells transfected with a plasmid encoding mcherry (cos) were added as a control for unbound mcherry protein detection with the anti-mcherry antibody ( arrow head ). c G protein activation: similar [ 35 S] GTPγS incorporation was measured on brain membranes from wild-type ( filled square ) ( n = 8), heterozygous ( n = 7) ( filled diamond ) and homozygous ( n = 11) ( filled circle ) mice following stimulation with the mu-selective agonist DAMGO. Data are the mean ± sem from independent experiments performed in triplicate ( n = 3 animals per genotype). d Tail immersion test: similar tail withdrawal latencies were measured at 52 °C in wild-type ( Oprm1 +/+ ) and MOR-mcherry ( Oprm1 mch/mch ) mice after saline or morphine injection (5 or 10 mg/kg, i.p.). Data are presented as mean ± sem ( n = 16 animals/group). * p
Figure Legend Snippet: Expression of functional receptors in MOR-mcherry knock-in mice. a Targeting strategy: Oprm1 exons, mcherry cDNA, and the FRT ( triangle ) flanked neomycin cassette are, respectively, displayed as exon number , mcherry , and neo . Homologous recombination (HR) was followed by FLP recombinase treatment (FLP) in ES cells. Positions of the oligonucleotides (BAZ 43, BAZ 44) used for genotyping are indicated. b Western blot: detection of MOR-mcherry fusion by immunoblotting with antibodies directed against mcherry on membranes from striatum and periaqueductal gray (PAG) from wild-type ( Oprm1 +/+ ), heterozygote ( Oprm1 +/mch ) and homozygote ( Oprm1 mch/mch ) mice (MOR-mcherry fusion indicated by arrow ). Cos cells transfected with a plasmid encoding mcherry (cos) were added as a control for unbound mcherry protein detection with the anti-mcherry antibody ( arrow head ). c G protein activation: similar [ 35 S] GTPγS incorporation was measured on brain membranes from wild-type ( filled square ) ( n = 8), heterozygous ( n = 7) ( filled diamond ) and homozygous ( n = 11) ( filled circle ) mice following stimulation with the mu-selective agonist DAMGO. Data are the mean ± sem from independent experiments performed in triplicate ( n = 3 animals per genotype). d Tail immersion test: similar tail withdrawal latencies were measured at 52 °C in wild-type ( Oprm1 +/+ ) and MOR-mcherry ( Oprm1 mch/mch ) mice after saline or morphine injection (5 or 10 mg/kg, i.p.). Data are presented as mean ± sem ( n = 16 animals/group). * p

Techniques Used: Expressing, Functional Assay, Knock-In, Mouse Assay, Homologous Recombination, Western Blot, Transfection, Plasmid Preparation, Activation Assay, Injection

5) Product Images from "pKa of opioid ligands as a discriminating factor for side effects"

Article Title: pKa of opioid ligands as a discriminating factor for side effects

Journal: Scientific Reports

doi: 10.1038/s41598-019-55886-1

Binding and activation of MOR. ( A ) Displacement of bound [³H]-DAMGO (4 nM) by FF6 at pH 6.5 and 7.4. ( B ) IC 50 calculated from ( A ). P > 0.05, unpaired t -test (n = 6–7). ( C ) [ 35 S]-GTPγS binding induced by FF6 at pH 6.5 and 7.4. [ 35 S]-GTPγS binding is expressed as percent increase in [ 35 S]-GTPγS binding relative to binding in unstimulated samples (n = 6). ( D ) EC 50 of FF6 from ( C ). P > 0.05, unpaired t -test. Data are presen t ed as mean ± SEM (A, C) and as mean ± 95% confidence intervals (B, D) (n = 6).
Figure Legend Snippet: Binding and activation of MOR. ( A ) Displacement of bound [³H]-DAMGO (4 nM) by FF6 at pH 6.5 and 7.4. ( B ) IC 50 calculated from ( A ). P > 0.05, unpaired t -test (n = 6–7). ( C ) [ 35 S]-GTPγS binding induced by FF6 at pH 6.5 and 7.4. [ 35 S]-GTPγS binding is expressed as percent increase in [ 35 S]-GTPγS binding relative to binding in unstimulated samples (n = 6). ( D ) EC 50 of FF6 from ( C ). P > 0.05, unpaired t -test. Data are presen t ed as mean ± SEM (A, C) and as mean ± 95% confidence intervals (B, D) (n = 6).

Techniques Used: Binding Assay, Activation Assay

6) Product Images from "Differential Modulation of ?- and ?-Opioid Receptor Agonists by Endogenous RGS4 Protein in SH-SY5Y Cells *"

Article Title: Differential Modulation of ?- and ?-Opioid Receptor Agonists by Endogenous RGS4 Protein in SH-SY5Y Cells *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M109.015453

Development of a SH-SY5Y cell line stably expressing shRNA against RGS4 . A , delivery of shRNA into SH-SY5Y cells. A mixture of four lentiviruses encoding four shRNAs targeting four different sites on the RGS4 gene was used to infect SH-SY5Y cells. A photograph using a Leica inverted fluorescence microscope at 5 days postinfection shows GFP expression as an indication of expressed shRNA in the cells ( left ) compared with the phase contrast ( right ). B , RGS4 protein knockdown. Whole cell lysates were prepared from the SH-SY5Y cells stably expressing shRNA against RGS4 or against GFP as a negative control, respectively. Equal amounts of protein (50 μg each) were loaded onto a 12% SDS-PAGE mini gel and blotted with anti-RGS4 antibody (U1079) (1:10,000 dilution), as described under “Experimental Procedures.” There was almost complete knockdown of RGS4 expression in cells expressing shRNA against RGS4 compared with cells expressing shRNA against GFP. The β-actin loading control was unchanged. C , expressed μ- and δ-opioid receptor numbers ( n = 3 in duplicate, mean ± S.E.) were the same in membranes from SH-SY5Y cells expressing shRNA against RGS4 or GFP determined with [ 3 H]DAMGO (μ) or [ 3 H]DPDPE (δ), as described under “Experimental Procedures.”
Figure Legend Snippet: Development of a SH-SY5Y cell line stably expressing shRNA against RGS4 . A , delivery of shRNA into SH-SY5Y cells. A mixture of four lentiviruses encoding four shRNAs targeting four different sites on the RGS4 gene was used to infect SH-SY5Y cells. A photograph using a Leica inverted fluorescence microscope at 5 days postinfection shows GFP expression as an indication of expressed shRNA in the cells ( left ) compared with the phase contrast ( right ). B , RGS4 protein knockdown. Whole cell lysates were prepared from the SH-SY5Y cells stably expressing shRNA against RGS4 or against GFP as a negative control, respectively. Equal amounts of protein (50 μg each) were loaded onto a 12% SDS-PAGE mini gel and blotted with anti-RGS4 antibody (U1079) (1:10,000 dilution), as described under “Experimental Procedures.” There was almost complete knockdown of RGS4 expression in cells expressing shRNA against RGS4 compared with cells expressing shRNA against GFP. The β-actin loading control was unchanged. C , expressed μ- and δ-opioid receptor numbers ( n = 3 in duplicate, mean ± S.E.) were the same in membranes from SH-SY5Y cells expressing shRNA against RGS4 or GFP determined with [ 3 H]DAMGO (μ) or [ 3 H]DPDPE (δ), as described under “Experimental Procedures.”

Techniques Used: Stable Transfection, Expressing, shRNA, Fluorescence, Microscopy, Negative Control, SDS Page

7) Product Images from "Modulation of μ‐opioid receptor activation by acidic pH is dependent on ligand structure and an ionizable amino acid residue. Modulation of μ‐opioid receptor activation by acidic pH is dependent on ligand structure and an ionizable amino acid residue"

Article Title: Modulation of μ‐opioid receptor activation by acidic pH is dependent on ligand structure and an ionizable amino acid residue. Modulation of μ‐opioid receptor activation by acidic pH is dependent on ligand structure and an ionizable amino acid residue

Journal: British Journal of Pharmacology

doi: 10.1111/bph.14810

Dependence of [ 35 S]‐GTPγS binding on pH and H297 6.52 . (a) Fentanyl‐ and (b) DAMGO‐induced [ 35 S]‐GTPγS binding to MOR‐WT and in MOR‐H297 6.52 A. Data represent mean ± SEM of specific [ 35 S]‐GTPγS binding in % of baseline with non‐linear fit. n = 6 per curve. Derived parameters are shown in Table 2 . (a) No significant differences were found; (b) significant differences ( P
Figure Legend Snippet: Dependence of [ 35 S]‐GTPγS binding on pH and H297 6.52 . (a) Fentanyl‐ and (b) DAMGO‐induced [ 35 S]‐GTPγS binding to MOR‐WT and in MOR‐H297 6.52 A. Data represent mean ± SEM of specific [ 35 S]‐GTPγS binding in % of baseline with non‐linear fit. n = 6 per curve. Derived parameters are shown in Table 2 . (a) No significant differences were found; (b) significant differences ( P

Techniques Used: Binding Assay, Derivative Assay

Related Articles

other:

Article Title: G Protein-Coupled Receptor Heteromerization: A Role in Allosteric Modulation of Ligand Binding S⃞
Article Snippet: [3 H]DAMGO and [3 H]deltorphin II were from PerkinElmer Life and Analytical Sciences (Waltham, MA).

Article Title: Mu-Opioid Receptor Coupling to G?o Plays an Important Role in Opioid Antinociception
Article Snippet: [3 H]DPN, [3 H]DAMGO and [35 S]GTPγS were purchased from PerkinElmer.

Article Title: Modulation of μ‐opioid receptor activation by acidic pH is dependent on ligand structure and an ionizable amino acid residue. Modulation of μ‐opioid receptor activation by acidic pH is dependent on ligand structure and an ionizable amino acid residue
Article Snippet: [3 H]‐naloxone, [3 H]‐DAMGO, and [35 S]‐GTPγS were purchased from Perkin Elmer (Waltham, USA).

Activity Assay:

Article Title: Dopamine D4 Receptor Counteracts Morphine-Induced Changes in ? Opioid Receptor Signaling in the Striosomes of the Rat Caudate Putamen
Article Snippet: .. Autoradiographic saturation kinetic study of MOR was performed using [3 H]DAMGO as radioligand (specific activity 56 Ci/mmol; PerkinElmer, Waltham, MA, USA). .. The sections were pre-incubated for 30 min at RT with 50 mM Tris-HCl (pH 7.4) and 5% BSA to remove endogenous opioids, and then incubated for 1 h with the same buffer containing [3 H]DAMGO (concentration ranging from 0.36 nM to 4 nM).

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    PerkinElmer radioligand saturation binding assay tritium tagged damgo
    <t>Radioligand</t> binding of rat and NMR MOR. A ) <t>[H3]DAMGO</t> binding to plasma membrane extracts of rat and NMR MOR expressed in HEK293 cells (pCMV- oprm1 -IRES-eGFP vector DNAs). Data are shown as whisker blots (min to max). B ) Normalized [H3]DAMGO saturation binding curve of rat and NMR MOR expressed in HEK293 cells (pCMV- oprm1 -IRES-eGFP vector DNAs) using whole membrane preparations. Data represent means ± SEM. Statistical analysis was performed using the two-tailed Mann Whitney U test.
    Radioligand Saturation Binding Assay Tritium Tagged Damgo, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/radioligand saturation binding assay tritium tagged damgo/product/PerkinElmer
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    radioligand saturation binding assay tritium tagged damgo - by Bioz Stars, 2020-09
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    92
    PerkinElmer damgo
    Association kinetics of δOR radioligand agonist binding. SK-N-SH whole cells endogenously expressing μOR and δOR (2:1 ratio) were incubated with 6 nM [ 3 <t>H]deltorphin</t> II in the absence or presence of 10 nM <t>DAMGO</t> or fentanyl for different
    Damgo, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/damgo/product/PerkinElmer
    Average 92 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    damgo - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    85
    PerkinElmer mu opioid receptor agonist damgo
    Mu <t>opioid</t> agonist-induced [ 35 S]-GTPγS binding is comparable on spinal cord membrane preparation from mu-cKO and mu fl mice. Spinal cord membranes were incubated in the absence or presence of the mu opioid agonist <t>DAMGO</t> (10 -9 -10 -4 M) in assay buffer containing [ 35 S] GTPγS. Basal level (100%) represents the specific [ 35 S]-GTPγS binding in the absence of agonist. DAMGO significantly increases [ 35 S]-GTPγS binding, in a comparable manner for mu-cKO and mu fl mice. [ 35 S]-GTPγS binding was absent on spinal cord membranes from mu-KO mice, and was decreased by half with a 50%-50% mix of membranes from mu fl and mu-KO animals, containing half mu receptors as compared to mu fl membranes, indicating that this assay allows to detect reduced receptor expression. Results are presented as means ± sem of 5-6 experiments on 5 distinct membrane preparations. ★★★ P
    Mu Opioid Receptor Agonist Damgo, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer h damgo
    Characterization of the binding properties of [3H] <t>DAMGO</t> and [ 3 H]‐Naloxone <t>(NLX)</t> to MOR expressed in HEK 293 cells and MOR expressed in a cell‐free synthesis system. (A) [ 3 H]‐DAMGO saturation binding experiments of HEK MOR and TCF‐MOR in presence of different detergents. (B and C) [ 3 H]‐NLX saturation binding experiments of HEK MOR and TCF‐MOR in the presence of different detergents. (D) [ 3 H]‐DAMGO single point binding experiment of HEK MOR, TCF‐MOR, NCF‐MOR, and NCMF‐MOR ( n = 6) presented as counts per minute (cpm). (E) [ 3 H]‐DAMGO single point binding experiment of HEK MOR, TCF‐MOR, NCFMOR, and NCMF‐MOR ( n = 6) presented as fmol/mg total protein.
    H Damgo, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 89/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Radioligand binding of rat and NMR MOR. A ) [H3]DAMGO binding to plasma membrane extracts of rat and NMR MOR expressed in HEK293 cells (pCMV- oprm1 -IRES-eGFP vector DNAs). Data are shown as whisker blots (min to max). B ) Normalized [H3]DAMGO saturation binding curve of rat and NMR MOR expressed in HEK293 cells (pCMV- oprm1 -IRES-eGFP vector DNAs) using whole membrane preparations. Data represent means ± SEM. Statistical analysis was performed using the two-tailed Mann Whitney U test.

    Journal: PLoS ONE

    Article Title: Functional Characteristics of the Naked Mole Rat ?-Opioid Receptor

    doi: 10.1371/journal.pone.0079121

    Figure Lengend Snippet: Radioligand binding of rat and NMR MOR. A ) [H3]DAMGO binding to plasma membrane extracts of rat and NMR MOR expressed in HEK293 cells (pCMV- oprm1 -IRES-eGFP vector DNAs). Data are shown as whisker blots (min to max). B ) Normalized [H3]DAMGO saturation binding curve of rat and NMR MOR expressed in HEK293 cells (pCMV- oprm1 -IRES-eGFP vector DNAs) using whole membrane preparations. Data represent means ± SEM. Statistical analysis was performed using the two-tailed Mann Whitney U test.

    Article Snippet: Radioligand saturation binding assay Tritium-tagged DAMGO ([D-Ala2, N-MePhe4, Gly-ol]-enkephalin, Perkin Elmer) was used as previously described [ ].

    Techniques: Binding Assay, Nuclear Magnetic Resonance, Plasmid Preparation, Whisker Assay, Two Tailed Test, MANN-WHITNEY

    Association kinetics of δOR radioligand agonist binding. SK-N-SH whole cells endogenously expressing μOR and δOR (2:1 ratio) were incubated with 6 nM [ 3 H]deltorphin II in the absence or presence of 10 nM DAMGO or fentanyl for different

    Journal: Molecular Pharmacology

    Article Title: G Protein-Coupled Receptor Heteromerization: A Role in Allosteric Modulation of Ligand Binding S⃞

    doi: 10.1124/mol.110.070847

    Figure Lengend Snippet: Association kinetics of δOR radioligand agonist binding. SK-N-SH whole cells endogenously expressing μOR and δOR (2:1 ratio) were incubated with 6 nM [ 3 H]deltorphin II in the absence or presence of 10 nM DAMGO or fentanyl for different

    Article Snippet: [3 H]DAMGO and [3 H]deltorphin II were from PerkinElmer Life and Analytical Sciences (Waltham, MA).

    Techniques: Binding Assay, Expressing, Incubation

    Dissociation kinetics of [ 3 H]deltorphin II (A) or [ 3 H]DAMGO (B) binding. A, SK-N-SH whole cells endogenously expressing μOR and δOR were incubated with 6 nM [ 3 H]deltorphin II for 1 h at 37°C. The supernatant was removed, the plates

    Journal: Molecular Pharmacology

    Article Title: G Protein-Coupled Receptor Heteromerization: A Role in Allosteric Modulation of Ligand Binding S⃞

    doi: 10.1124/mol.110.070847

    Figure Lengend Snippet: Dissociation kinetics of [ 3 H]deltorphin II (A) or [ 3 H]DAMGO (B) binding. A, SK-N-SH whole cells endogenously expressing μOR and δOR were incubated with 6 nM [ 3 H]deltorphin II for 1 h at 37°C. The supernatant was removed, the plates

    Article Snippet: [3 H]DAMGO and [3 H]deltorphin II were from PerkinElmer Life and Analytical Sciences (Waltham, MA).

    Techniques: Binding Assay, Expressing, Incubation

    Mu opioid agonist-induced [ 35 S]-GTPγS binding is comparable on spinal cord membrane preparation from mu-cKO and mu fl mice. Spinal cord membranes were incubated in the absence or presence of the mu opioid agonist DAMGO (10 -9 -10 -4 M) in assay buffer containing [ 35 S] GTPγS. Basal level (100%) represents the specific [ 35 S]-GTPγS binding in the absence of agonist. DAMGO significantly increases [ 35 S]-GTPγS binding, in a comparable manner for mu-cKO and mu fl mice. [ 35 S]-GTPγS binding was absent on spinal cord membranes from mu-KO mice, and was decreased by half with a 50%-50% mix of membranes from mu fl and mu-KO animals, containing half mu receptors as compared to mu fl membranes, indicating that this assay allows to detect reduced receptor expression. Results are presented as means ± sem of 5-6 experiments on 5 distinct membrane preparations. ★★★ P

    Journal: PLoS ONE

    Article Title: Mu Opioid Receptors on Primary Afferent Nav1.8 Neurons Contribute to Opiate-Induced Analgesia: Insight from Conditional Knockout Mice

    doi: 10.1371/journal.pone.0074706

    Figure Lengend Snippet: Mu opioid agonist-induced [ 35 S]-GTPγS binding is comparable on spinal cord membrane preparation from mu-cKO and mu fl mice. Spinal cord membranes were incubated in the absence or presence of the mu opioid agonist DAMGO (10 -9 -10 -4 M) in assay buffer containing [ 35 S] GTPγS. Basal level (100%) represents the specific [ 35 S]-GTPγS binding in the absence of agonist. DAMGO significantly increases [ 35 S]-GTPγS binding, in a comparable manner for mu-cKO and mu fl mice. [ 35 S]-GTPγS binding was absent on spinal cord membranes from mu-KO mice, and was decreased by half with a 50%-50% mix of membranes from mu fl and mu-KO animals, containing half mu receptors as compared to mu fl membranes, indicating that this assay allows to detect reduced receptor expression. Results are presented as means ± sem of 5-6 experiments on 5 distinct membrane preparations. ★★★ P

    Article Snippet: Samples were incubated in triplicate with and without the mu opioid receptor agonist DAMGO (10-9 to 10-4 M), for 1 hour at 25°C in assay buffer containing 30 µM GDP and 0.1nM [35 S] GTPγS (NEG030H PerkinElmer).

    Techniques: Binding Assay, Mouse Assay, Incubation, Expressing

    Characterization of the binding properties of [3H] DAMGO and [ 3 H]‐Naloxone (NLX) to MOR expressed in HEK 293 cells and MOR expressed in a cell‐free synthesis system. (A) [ 3 H]‐DAMGO saturation binding experiments of HEK MOR and TCF‐MOR in presence of different detergents. (B and C) [ 3 H]‐NLX saturation binding experiments of HEK MOR and TCF‐MOR in the presence of different detergents. (D) [ 3 H]‐DAMGO single point binding experiment of HEK MOR, TCF‐MOR, NCF‐MOR, and NCMF‐MOR ( n = 6) presented as counts per minute (cpm). (E) [ 3 H]‐DAMGO single point binding experiment of HEK MOR, TCF‐MOR, NCFMOR, and NCMF‐MOR ( n = 6) presented as fmol/mg total protein.

    Journal: Biotechnology and Bioengineering

    Article Title: Production of G protein‐coupled receptors in an insect‐based cell‐free system

    doi: 10.1002/bit.26346

    Figure Lengend Snippet: Characterization of the binding properties of [3H] DAMGO and [ 3 H]‐Naloxone (NLX) to MOR expressed in HEK 293 cells and MOR expressed in a cell‐free synthesis system. (A) [ 3 H]‐DAMGO saturation binding experiments of HEK MOR and TCF‐MOR in presence of different detergents. (B and C) [ 3 H]‐NLX saturation binding experiments of HEK MOR and TCF‐MOR in the presence of different detergents. (D) [ 3 H]‐DAMGO single point binding experiment of HEK MOR, TCF‐MOR, NCF‐MOR, and NCMF‐MOR ( n = 6) presented as counts per minute (cpm). (E) [ 3 H]‐DAMGO single point binding experiment of HEK MOR, TCF‐MOR, NCFMOR, and NCMF‐MOR ( n = 6) presented as fmol/mg total protein.

    Article Snippet: Briefly, 100 μg of cell membranes were prepared and incubated for 90 min in assay buffer (50 mM Trizma, pH 7.4) with increasing doses of [3 H]‐DAMGO (0.5–16 nM) (47.1 Ci/mmol) and [3 H]‐NLX (0.9–15 nM) (58.2 Ci/mmol) (Perkin Elmer, Waltham, MA), respectively, for saturation binding studies or with 4 nM [3 H]‐DAMGO for single point binding studies in the absence or presence of 10 μM unlabeled NLX to determine nonspecific binding.

    Techniques: Binding Assay