Structured Review

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

Journal: Biotechnology and Bioengineering

doi: 10.1002/bit.26346

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.
Figure Legend 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.

Techniques Used: Binding Assay

Related Articles

Incubation:

Article Title: Antinociception produced by 14,15-epoxyeicosatrienoic acid is mediated by the activation of ?-endorphin and Met-enkephalin in the rat ventrolateral periaqueductal gray
Article Snippet: .. The membrane homogenate (900–1000 μg protein/assay) was incubated at 25°C for 2 h in 50 mM Tris–HCl buffer (pH 7.4) with 1 nM [3 H]DAMGO (specific activity, 56.8 Ci/mmol) or 5nM [3 H]Naltrindole (specific activity, 20.0 Ci/mmol, PerkinElmer Life and Analytical Sciences, Boston, MA) in a total volume of 1 ml. .. The reaction was terminated using a Brandel cell harvester (Model M-24, Brandel, MD) and the samples were filtered through Whatman GF/B glass filters pre-soaked in 50 mM Tris–HCl (pH 7.4) with 0.01% polyethylenimine at 4°C for 2 h. Filters were washed three times with 5 ml of Tris–HCl buffer (pH 7.4, 4°C) and then transferred to scintillation counting vials containing 0.5 ml of a tissue solubilizer (Soluene-350, Packard Instrument Company, Meriden, CT) and 4 ml of a scintillation cocktail (Hionic Fluor, Packard Instrument Company).

Article Title: Production of G protein‐coupled receptors in an insect‐based cell‐free system
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. .. The conversion of counts per min (cpm) to fmol/mg total protein was realized using the following equation: (cpm × 100)/(counter efficacy × 2.2 × specific activity of [3 H]‐DAMGO (or [3 H]‐NLX) × total protein amount in mg).

Binding Assay:

Article Title: Neocosmospora sp-derived resorcylic acid lactones with in vitro binding affinity for human opioid and cannabinoid receptors
Article Snippet: .. All chemicals used were purchased from Sigma-Aldrich (Poole, Dorset, U.K.) with the following exceptions: for the binding experiments, [3 H]-CP-55,940 (174.8 Ci/mmol), [3 H]-DAMGO (53.4 Ci/mmol), [3 H]-U-69,593 (42.7 Ci/mmol), and [3 H]-enkephalin (45 Ci/mmol) were purchased from Perkin-Elmer Life Sciences Inc. (Boston, MA, U.S.A.). .. CP-55,940, DAMGO, DPDPE, nor-binaltorphimine, and WIN 55,212-2 mesylate were purchased from Tocris Bioscience (Ellisville, Missouri, U.S.A.).

Article Title: Wheel running during chronic nicotine exposure is protective against mecamylamine‐precipitated withdrawal and up‐regulates hippocampal α7 nACh receptors in mice
Article Snippet: .. [125 I]‐epibatidine (specific activity 2200 Ci·mmol−1 ), [125 I]‐α‐bungarotoxin (specific activity 108.8 Ci·mmol−1 ), [3 H]‐DAMGO (specific activity 51.5 Ci·mmol−1 ) and [3 H]‐raclopride (specific activity 60 Ci·mmol−1 ) used for autoradiographic binding experiments were purchased from PerkinElmer (Waltham, MA, USA). ..

Article Title: Production of G protein‐coupled receptors in an insect‐based cell‐free system
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. .. The conversion of counts per min (cpm) to fmol/mg total protein was realized using the following equation: (cpm × 100)/(counter efficacy × 2.2 × specific activity of [3 H]‐DAMGO (or [3 H]‐NLX) × total protein amount in mg).

Activity Assay:

Article Title: Wheel running during chronic nicotine exposure is protective against mecamylamine‐precipitated withdrawal and up‐regulates hippocampal α7 nACh receptors in mice
Article Snippet: .. [125 I]‐epibatidine (specific activity 2200 Ci·mmol−1 ), [125 I]‐α‐bungarotoxin (specific activity 108.8 Ci·mmol−1 ), [3 H]‐DAMGO (specific activity 51.5 Ci·mmol−1 ) and [3 H]‐raclopride (specific activity 60 Ci·mmol−1 ) used for autoradiographic binding experiments were purchased from PerkinElmer (Waltham, MA, USA). ..

Article Title: Antinociception produced by 14,15-epoxyeicosatrienoic acid is mediated by the activation of ?-endorphin and Met-enkephalin in the rat ventrolateral periaqueductal gray
Article Snippet: .. The membrane homogenate (900–1000 μg protein/assay) was incubated at 25°C for 2 h in 50 mM Tris–HCl buffer (pH 7.4) with 1 nM [3 H]DAMGO (specific activity, 56.8 Ci/mmol) or 5nM [3 H]Naltrindole (specific activity, 20.0 Ci/mmol, PerkinElmer Life and Analytical Sciences, Boston, MA) in a total volume of 1 ml. .. The reaction was terminated using a Brandel cell harvester (Model M-24, Brandel, MD) and the samples were filtered through Whatman GF/B glass filters pre-soaked in 50 mM Tris–HCl (pH 7.4) with 0.01% polyethylenimine at 4°C for 2 h. Filters were washed three times with 5 ml of Tris–HCl buffer (pH 7.4, 4°C) and then transferred to scintillation counting vials containing 0.5 ml of a tissue solubilizer (Soluene-350, Packard Instrument Company, Meriden, CT) and 4 ml of a scintillation cocktail (Hionic Fluor, Packard Instrument Company).

other:

Article Title: Differential Effect of Membrane Cholesterol Removal on ?- and ?-Opioid Receptors
Article Snippet: [3 H]Diprenorphine, [3 H]DAMGO ([ d -Ala2 , N -Me-Phe4 ,Gly5 -ol]enkephalin), and [35 S]GTPγS were obtained from PerkinElmer Life Sciences.

Article Title: Pharmacological Characterization of Dezocine, a Potent Analgesic Acting as a κ Partial Agonist and μ Partial Agonist
Article Snippet: [3 H]DAMGO (51.5 Ci/mmol), [3 H]DPDPE (57.4 Ci/mmol), [3 H]U69593 (43.6 Ci/mmol) and [35 S]GTPγS (1250 Ci/mmol) were bought from Perkin Elmer.

Article Title: REDUCED EXPRESSION OF THE MU OPIOID RECEPTOR IN SOME, BUT NOT ALL, BRAIN REGIONS IN MICE WITH Oprm1 A112G
Article Snippet: [3 H]DAMGO (56.0 Ci/mmol) and tritium-sensitive storage phosphor screens were purchased from Perkin Elmer (Boston, MA, USA).

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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
    https://www.bioz.com/result/h damgo/product/PerkinElmer
    Average 89 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    h damgo - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

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    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

    Opioid receptor antagonist and agonist modulation MOR- or DOR-specific ligand binding in NK cells. Shown are representative saturation curves ( A, C, E, and G ) and the B max ( B, D, F, and H ) of MOR-specific ligand [ 3 H]DAMGO or DOR-specific ligand [ 3 H]naltrindole.

    Journal: The Journal of Biological Chemistry

    Article Title: Opiate Antagonist Prevents ?- and ?-Opiate Receptor Dimerization to Facilitate Ability of Agonist to Control Ethanol-altered Natural Killer Cell Functions and Mammary Tumor Growth *

    doi: 10.1074/jbc.M112.347583

    Figure Lengend Snippet: Opioid receptor antagonist and agonist modulation MOR- or DOR-specific ligand binding in NK cells. Shown are representative saturation curves ( A, C, E, and G ) and the B max ( B, D, F, and H ) of MOR-specific ligand [ 3 H]DAMGO or DOR-specific ligand [ 3 H]naltrindole.

    Article Snippet: Spleens of experimental animals were dissociated, and erythrocytes were removed by a 5-s hypotonic shock with sterile distilled water, and splenocytes were isolated and incubated at a concentration of 1–4 × 106 /well with various concentrations of [3 H]naltrindole or [3 H]DAMGO (PerkinElmer Life Sciences) in 96-well plates for 4.5 h at room temperature in Ca2+ - and Mg2+ -deficient Hanks' balanced salt solution.

    Techniques: Ligand Binding Assay

    Imposition of fentanyl (red, our simulations) and DAMGO (yellow, PDB structure: 6DDF [ 53 ]) in μOR (blue), with focus on similar positioning of fentanyl anilide’s phenyl and phenyl of N-Me-Phe 4 residue of DAMGO.

    Journal: Molecules

    Article Title: Fentanyl Family at the Mu-Opioid Receptor: Uniform Assessment of Binding and Computational Analysis

    doi: 10.3390/molecules24040740

    Figure Lengend Snippet: Imposition of fentanyl (red, our simulations) and DAMGO (yellow, PDB structure: 6DDF [ 53 ]) in μOR (blue), with focus on similar positioning of fentanyl anilide’s phenyl and phenyl of N-Me-Phe 4 residue of DAMGO.

    Article Snippet: The so-obtained membrane fractions (containing opioid receptors) were incubated at 25 °C for 60 min in the presence of 0.5 nM [3 H]DAMGO (a radioligand specific for μOR) bought from PerkinElmer, USA, and the increasing concentrations of the assayed compounds (3.0 × 10−10 up to 10−5 M, each concentration in duplicate).

    Techniques:

    Saturation binding curve of [ 3 H]-DAMGO in intact CHO-MOPr-WT and CHO-MOPr-N40D cells, 24 h after induction of receptor expression with tetracycline. Radioligand binding was carried out as described in the Methods. No significant difference in B max or

    Journal: British Journal of Pharmacology

    Article Title: Buprenorphine signalling is compromised at the N40D polymorphism of the human μ opioid receptor in vitro

    doi: 10.1111/bph.12785

    Figure Lengend Snippet: Saturation binding curve of [ 3 H]-DAMGO in intact CHO-MOPr-WT and CHO-MOPr-N40D cells, 24 h after induction of receptor expression with tetracycline. Radioligand binding was carried out as described in the Methods. No significant difference in B max or

    Article Snippet: Surface expression of MOP receptors was determined on intact CHO-MOP receptor cells by incubation with 0.125–16 nM [3 H]-DAMGO (D-Ala2 , N-MePhe4 , Gly-ol]-enkephalin; PerkinElmer, Waltham, MA, USA) at 4°C in 50 mM Tris-Cl (pH 7.4) for 2 h. Briefly, 24 h before the assay, cells were detached from flasks with trypsin/EDTA (Sigma, Castle Hill, NSW, Australia) and resuspended in DMEM containing 10% FBS, 100 U penicillin/streptomycin, plus 2 μg·mL−1 tetracycline to induce MOP receptor expression.

    Techniques: Binding Assay, Expressing

    DAMGO inhibits AC and activates ERK1/2 in CHO cells expressing MOPr-WT or MOPr-N40D. AC inhibition and levels of ERK1/2 phosphorylation were determined as described in the Methods. (A) Traces showing changes in fluorescent signal following application

    Journal: British Journal of Pharmacology

    Article Title: Buprenorphine signalling is compromised at the N40D polymorphism of the human μ opioid receptor in vitro

    doi: 10.1111/bph.12785

    Figure Lengend Snippet: DAMGO inhibits AC and activates ERK1/2 in CHO cells expressing MOPr-WT or MOPr-N40D. AC inhibition and levels of ERK1/2 phosphorylation were determined as described in the Methods. (A) Traces showing changes in fluorescent signal following application

    Article Snippet: Surface expression of MOP receptors was determined on intact CHO-MOP receptor cells by incubation with 0.125–16 nM [3 H]-DAMGO (D-Ala2 , N-MePhe4 , Gly-ol]-enkephalin; PerkinElmer, Waltham, MA, USA) at 4°C in 50 mM Tris-Cl (pH 7.4) for 2 h. Briefly, 24 h before the assay, cells were detached from flasks with trypsin/EDTA (Sigma, Castle Hill, NSW, Australia) and resuspended in DMEM containing 10% FBS, 100 U penicillin/streptomycin, plus 2 μg·mL−1 tetracycline to induce MOP receptor expression.

    Techniques: Expressing, Inhibition