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CP-55,940

CP-55,940 is a synthetic cannabinoid receptor agonist with potent psychoactive effects.
It is used extensively in research to study the endocannabinoid system and its role in various physiological and pathological processes.
PubCompare.ai can enhance your CP-55,940 research by helping you locate relevant protocols from literature, pre-prints, and patents, and leveraging AI-driven comparisons to identify the best protocols and products.
This can improve your reproducibility and accuracy, leading to more robust and reliable findings.

Most cited protocols related to «CP-55,940»

Forebrain synaptosomal membranes were prepared from frozen rat brains by the method described by Dodd et al.36 (link) and were used to assess the affinities of the novel analogs for the CB1 binding sites, while affinities for the CB2 sites were measured using a membrane preparation from frozen mouse spleen using a similar procedure.37 (link) Membrane preparations from HEK293 cells expressing human CB2 (hCB2) receptor were used to assess the affinities of representative analogs for hCB2.33 (link) The displacement of specifically tritiated CP-55,940 from these membranes was used to determine the IC50 values for the test compounds. The assay was conducted in a 96-well microfilter plate. The samples were filtered using a Packard Filtermate Harvester and Whatman GF/B unifilter-96 plates, and 0.5% BSA was incorporated into the wash buffer. Radioactivity was detected using MicroScint 20 scintillation cocktail added to the dried filter plates and was counted using a Packard Instruments Top Count. Data were collected from three independent experiments between 100% and 0% specific binding for [3H]CP-55,940, determined using 0 and 100 nM CP-55,940. The normalized data from three independent experiments were combined and analyzed using a four-parameter logistic equation to yield IC50 values that were converted to Ki values using the assumptions of Cheng and Prussoff.38 (link)
Publication 2010
Binding Sites Biological Assay Brain Buffers CP-55,940 Freezing Homo sapiens Mice, House Plasma Membrane Prosencephalon Radioactivity Receptor, Cannabinoid, CB2 Spleen Synaptosomes Tissue, Membrane
CHO-K1 hCB1R and hCB2R cells were disrupted by cavitation in a pressure cell, and membranes were sedimented by ultracentrifugation, as described by Bolognini et al.34 (link),58 (link). The pellet was resuspended in TME buffer (50 mM Tris–HCl, 5 mM MgCl2, 1 mM EDTA, pH 7.4) and membrane proteins were quantified with a Bradford dye-binding method (Bio-Rad Laboratories, Mississauga, ON).
Assays were carried out with [3H]CP55,940 and Tris binding buffer (50 mM Tris–HCl, 50 mM Tris–base, 0.1% BSA, pH 7.4), total assay volume 2 mL, using the filtration procedure described previously by Baillie et al.18 (link). The binding was initiated by the addition of transfected human CHO-K1 hCB1R and hCB2R cell membranes (50 µg protein per well). All assays were performed at 37 °C for 60 min before termination by the addition of ice-cold Tris binding buffer, followed by vacuum filtration using a 24-well sampling manifold (Brandel Cell Harvester; Brandel Inc, Gaithersburg, MD, USA) and Brandel GF/B filters that had been soaked in wash buffer at 4 °C for at least 24 h. Each reaction well was washed 6 times with 1.2 mL aliquots of Tris-binding buffer. The filters were air-dried overnight and then placed in 5 mL of scintillation fluid (Ultima Gold XR, PerkinElmer). Radioactivity was quantified by liquid scintillation spectrometry. Specific binding was defined as the difference between the binding that occurred in the presence and absence of 1 µM unlabelled CP55,940. The concentration of [3H]CP55940 used in our displacement assays was 0.7 nM.
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Publication 2020
Biological Assay Buffers Cells Cold Temperature CP-55,940 Edetic Acid Filtration Gold Homo sapiens Magnesium Chloride Membrane Proteins Plasma Membrane Pressure Proteins Radioactivity Spectrometry Tissue, Membrane Tromethamine Ultracentrifugation Vacuum
Each compound was pharmacologically characterized using a functional fluorescent CB1 activated Gαq16-coupled intracellular calcium mobilization assay in CHO-K1 cells as has been described in our previous publications and apparent affinity (Ke) values were determined.7 (link) Briefly, CHO-K1 cells were engineered to co-express human CB1 and Gqα16. Activation of CB1 by an agonist then leads to generation of inositol phospahatase 3 (IP3) and activation of IP3 receptors, which leads to mobilization of intracellular calcium. Calcium flux was monitored in a 96-well format using the fluorescent dye Calcein-4 AM in an automated platereader (Flexstation, Molecular Devices). The antagonism of a test compound was measured by its ability to shift the concentration response curve of the synthetic CB1 agonist CP55940 rightwards using the equation:
Ke=[Ligand]/[DR1] where DR is the EC50 ratio of CP55940 in the presence or absence of a test agent.14 Further characterization of select compounds was performed using radioligand displacement of [3H]SR141716 and equilibrium dissociation constant (Ki) values were determined as has been described previously.7 (link), 15 (link) Selectivity of these compounds at CB1 versus CB2 was also determined by obtaining Ki values at either receptor using displacement of [3H]CP55940 in membranes of CHO-K1cells over-expressing either receptor. Data reported are average values from 3–6 measurements.
Publication 2012
antagonists Biological Assay Calcium CHO Cells CP-55,940 Fluorescent Dyes fluorexon Genetic Selection Homo sapiens Inositol Ligands Medical Devices Protoplasm SR141716 Tissue, Membrane
Each compound was pharmacologically characterized using a functional fluorescent CB1 activated Gαq16-coupled intracellular calcium mobilization assay in CHO-K1 cells, as has been described in our previous publications and apparent affinity (Ke) values were determined.13 (link) Briefly, CHO-K1 cells were engineered to coexpress human CB1 and Gαq16. Activation of CB1 by an agonist then leads to generation of inositol phospahatase 3 (IP3) and activation of IP3 receptors, which leads to mobilization of intracellular calcium. Calcium flux was monitored in a 96-well format using the fluorescent dye Calcein-4 a.m. in an automated platereader (Flexstation, Molecular Devices). The antagonism of a test compound was measured by its ability to shift the concentration response curve of the synthetic CB1 agonist CP55940 rightwards using the equation:
Ke = [Ligand]/[DR-1] where DR is the EC50 ratio of CP55940 in the presence or absence of a test agent. Standard errors were between 5 and 25% of mean in most cases and have been left out for clarity.
Further characterization of select compounds was performed using radioligand displacement of [3H]SR141716 and equilibrium dissociation constant (Ki) values were determined as described previously.13 (link) Selectivity of these compounds at CB1 versus CB2 was also determined by obtaining Ki values at either receptor using displacement of [3H]CP55940 in membranes of CHO-K1 cells overexpressing either receptor. Data reported are average values from 3 to 6 measurements. The standard errors for most measurements were between 5 and 25% of mean and have been left out from the tables and figures for clarity.
Publication 2012
antagonists Biological Assay Calcium CHO Cells CP-55,940 Fluorescent Dyes fluorexon Genetic Selection Homo sapiens Inositol Ligands Medical Devices Protoplasm SR141716 Tissue, Membrane

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Publication 2012
Brain Buffers Cannabinoids Cold Temperature CP-55,940 Ethanol Filtration Magnesium Chloride Mice, House Radioactivity Serum Albumin, Bovine Spectrophotometry Tromethamine Vacuum WIN 55,212

Most recents protocols related to «CP-55,940»

CP 55,940 (Merck Life Science Ltd., Gillingham, UK), CB1 and CB2 receptors agonist, CB1- and CB2- selective inverse agonists, SR 141716 (SR1) and SR 144528 (SR2) (Bio-Techne Ltd., Abingdon, UK), were dissolved in dimethyl sulfoxide (DMSO) and all stored at 10 mM stock solutions (−20 °C).
Appropriate concentrations were freshly prepared from stock solution using a culture medium. DMSO and ethanol diluent controls were also in. For binding experiments, [35S] guanosine 5′-O-[gamma-thio] triphosphate (GTPγS) (1250 Ci/mmol) was obtained from PerkinElmer Life Sciences (Stapeley, Nantwich, UK), GTPγS and adenosine deaminase from Roche Diagnostic (Merck Life Science Ltd., Gillingham, UK) and guanosine diphosphate (GDP) and phenylmethylsulfonyl fluoride (PMSF) from Merck Life Science Ltd., Gillingham, UK. All cells and chemicals were handled using the appropriate personal protective equipment. The DIM concentrations used in this study were chosen according to pharmacokinetic studies in animals and previous studies in prostate cancer cell lines [52 (link)].
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Publication 2023
agonists Animals Cell Lines Cells CP-55,940 Culture Media Deaminase, Adenosine Diagnosis Ethanol Gamma Rays Guanosine Guanosine Diphosphate Phenylmethylsulfonyl Fluoride Prostate Cancer Receptor, Cannabinoid, CB2 SR141716 SR 144528 Sulfoxide, Dimethyl triphosphate
Membrane preparations
from Chem-1 cells expressing the human CB1 receptors (ChemiSCREEN
CB1 Cannabinoid Receptor Membrane Preparation. Merck, USA) were incubated
in duplicate with 1 nM [3H]CP-55,940 (specific activity:
108.5 Ci/mmole, PerkinElmer, USA) in a 50 mM Tris–HCl, pH =
7.4 buffer supplemented with 1 mM CaCl2, 5 mM MgCl2, 0.2% BSA and increasing concentrations of the compounds
tested. Compounds were dissolved in 50% DMSO and added to the reaction
mixture at 10 concentrations equally spaced on a log scale (10–10–10–4.5 M). The final DMSO
concentration was 5%. Nonspecific binding was determined with 10 μM
WIN 55,212-2. The reaction mixture (500 μL) was incubated for
1.5 h at 30 °C. Before harvesting, Brandel Whatman GF/B Filter
Paper was presoaked with 0.5% polyethylenimine buffer for 30 min and
then washed with 2 mL of 50 mM Tris–HCl buffer (pH = 7.4) containing
0.5% BSA to minimize nonspecific binding. The reaction was terminated
by depositing the samples onto the filter paper with the Brandel M-24
Cell Harvester. Samples were then rapidly washed three times with
2 mL of wash buffer (50 mM Tris–HCl pH 7.4, 500 mM NaCl, and
5 mM MgCl2) to separate the bound radioligand from the
free one. The rest of the procedure was the same as for CB2.
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Publication 2023
Buffers CP-55,940 Homo sapiens Magnesium Chloride Plasma Membrane Polyethyleneimine Receptor, Cannabinoid Receptor, Cannabinoid, CB1 Sodium Chloride Sulfoxide, Dimethyl Tissue, Membrane Tromethamine
Ten micromolar
concentrations of each compound were incubated in triplicate with
membrane preparations from CHO-K1 cells expressing the human CB2 receptor
(0.5 μg per well) (PerkinElmer, Cat. No. ES-111-M400UA) in an
assay buffer containing 50 mM Tris–HCl, pH = 7.4, 0.2 mM EGTA,
3 mM MgCl2, 100 mM NaCl, 30 μM GDP and 1 mg/mL BSA)
in the presence of 0.08 nM [35S] guanosine 5′-[γ-thio]triphosphate
([35S]GTPγS) (specific activity: 1250 Ci/mmole, PerkinElmer).
Nonspecific binding was determined with 100 μM of unlabeled
GTPγS. CP-55,940 (100 nM) was used as stimulating ligand. The
final DMSO concentration in the assay was 5%. The reaction mixture
was incubated for 60 min at 30 °C. Next, the samples were deposited
under vacuum with the FilterMate Harvester (PerkinElmer, USA) onto
Unifilter GF/B Plates (PerkinElmer, USA) presoaked with wash buffer
(50 mM Tris–HCl, pH = 7.4). The samples were then rapidly washed
with 2 mL of wash buffer. Filter plates were dried for 30 min at 50
°C and 40 μL of MicroScint PS (PerkinElmer, USA) scintillation
fluid was added to each well. Radioactivity was counted in a Trilux
MicroBeta2 counter (PerkinElmer, USA). Data were analyzed
with GraphPad Prism 5.0 software. Results were expressed as percent
of basal [35S]GTPγS binding in the presence of CP-55,940
from three separate experiments. Basal binding was set to 100%.
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Publication 2023
Biological Assay Buffers CHO Cells CP-55,940 Egtazic Acid Guanosine Homo sapiens Ligands Magnesium Chloride prisma Radioactivity Receptor, Cannabinoid, CB2 Sodium Chloride Strains Sulfoxide, Dimethyl triphosphate Tromethamine Vacuum
The compounds selected
for the in vitro binding assay were purchased via MolPort, SIA, Riga,
Latvia (Supporting Information S2). Membrane
preparations from CHO-K1 cells expressing the human CB2 (ChemiSCREEN
Membrane Preparation Recombinant Human CB2 Cannabinoid Receptor. Merck,
USA) were incubated in duplicate with 0.8 nM [3H]CP-55,940
(specific activity: 101 Ci/mmole, PerkinElmer, USA) in a 50 mM Tris–HCl,
pH = 7.4 buffer supplemented with 2.5 mM EDTA, 5 mM MgCl2, 0.5 mg/mL BSA and increasing concentrations of the compounds tested.
Compounds were dissolved in 50% DMSO and added to the reaction mixture
at 10 concentrations equally spaced on a log scale (10–10–10–4.5 M). The final DMSO concentration
was 5%. Nonspecific binding was determined with 10 μM WIN 55,212-2.
The reaction mixture (500 μL) was incubated for 1.5 h at 30
°C. Before harvesting, Brandel Whatman GF/B Filter Paper was
presoaked with 0.5% polyethylenimine buffer for 30 min and then washed
with 2 mL of 50 mM Tris–HCl buffer (pH = 7.4) containing 0.5%
BSA to minimize nonspecific binding. The reaction was terminated by
depositing the samples onto the filter paper with the Brandel M–24
Cell Harvester. Samples were then rapidly washed three times with
2 mL of wash buffer (50 mM Tris–HCl pH 7.4, 2.5 mM EDTA, 5
mM MgCl2, 0.5 mg/mL BSA) to separate the bound radioligand
from free. Filters were then air-dried for 1.5 h at 60 °C. After
drying, filter discs were placed on a flexible 24-well plate and 500
μL of EcoScint-20 scintillant (PerkinElmer, USA) was added to
each well. Plates were counted (2 min per well) in a Trilux MicroBeta
counter (PerkinElmer, USA). Data were analyzed with GraphPad Prism
5.0 software. Curves were fitted with a one-site nonlinear regression
model, and inhibitory constants (pKi ±
SEM and Ki, 95% CI) were calculated from
the Cheng–Prusoff equation.
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Publication 2023
Biological Assay Buffers CHO Cells CP-55,940 Edetic Acid Homo sapiens Magnesium Chloride Polyethyleneimine Psychological Inhibition Receptor, Cannabinoid Sulfoxide, Dimethyl Tromethamine WIN 55,212
3 × 104 cells/well
were employed to seed cells onto 96-well plates, which were subsequently
incubated throughout the night. Cells were then treated with 2 μM
Fluo-4 AM in HBSS (0.3 mM Na2HPO4, 5.4 mM KCl,
4.2 mM NaHCO3, 0.4 mM KH2PO4, 0.5
mM MgCl2, 1.3 mM CaCl2, 137 mM NaCl, 0.6 mM
MgSO4, 250 μM sulfinpyrazone, and 5.6 mM d-glucose, pH 7.4) for 45 min at 37 °C. In agonist mode, the
extra dye was washed away and 50 μL of HBSS was included, followed
by dispensation of 25 μL of HBSS with the test compound, DMSO
(negative control), or CP55940 (positive control), into the wells
through a FlexStation microplate reader. Meanwhile, the change of
intracellular calcium was measured at an excitation wavelength of
525 and 485 nm. For antagonist mode, the extra dye was washed out
and 50 μL of HBSS with test compounds, DMSO (negative control),
or AM630 (positive control) was introduced. Following a 10 min incubation
under room temperature, 25 μL of CP55940 was placed in the wells
utilizing a FlexStation microplate reader. The change of intracellular
calcium was captured at an excitation wavelength of 525 and 485 nm.
To be noticed, each experiment has been independently replicated three
times, with three repetitions and eight gradients each time.
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Publication 2023
AM 630 Bicarbonate, Sodium Calcium Cells CP-55,940 Glucose Hemoglobin, Sickle Magnesium Chloride Sodium Chloride Sulfinpyrazone Sulfoxide, Dimethyl

Top products related to «CP-55,940»

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CP55,940 is a high-affinity cannabinoid receptor agonist. It binds to both CB1 and CB2 receptors with high potency.
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CP55940 is a synthetic cannabinoid receptor agonist. It functions as a potent and selective agonist for the CB1 and CB2 cannabinoid receptors.
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CP55,940 is a synthetic cannabinoid compound. It acts as a potent agonist at the CB1 and CB2 cannabinoid receptors. CP55,940 is commonly used in research to study the effects of cannabinoid receptor activation.
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AM251 is a synthetic cannabinoid receptor antagonist. It functions by selectively binding to and inhibiting the CB1 cannabinoid receptor.
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[35S]GTPγS is a radiolabeled GTP analog used in biochemical assays to study guanine nucleotide binding proteins and their interactions.
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More about "CP-55,940"

CP-55,940 is a synthetic cannabinoid receptor agonist, known for its potent psychoactive effects.
It has been extensively used in research to study the endocannabinoid system and its role in various physiological and pathological processes.
The compound can interact with both CB1 and CB2 receptors, making it a valuable tool for investigating the complex mechanisms underlying the endocannabinoid system.
In addition to CP-55,940, other related compounds like AM251, WIN55,212-2, and AM630 have also been utilized in endocannabinoid research.
These compounds, along with the radioligand [35S]GTPγS, have helped scientists understand the signaling pathways and pharmacological properties of the endocannabinoid system.
When conducting experiments with CP-55,940, researchers often employ solvents like DMSO to ensure proper dissolution and administration.
GF/C filters are commonly used to separate bound and unbound radioligands in receptor binding assays, providing insights into the receptor-ligand interactions.
By leveraging PubCompare.ai, researchers can enhance their CP-55,940 studies by accessing relevant protocols from literature, preprints, and patents.
The AI-driven comparisons offered by PubCompare.ai can help identify the most robust and reliable protocols, improving reproducibility and accuracy in their findings.
This can lead to more robust and reliable research outcomes, advancing our understanding of the endocannabinoid system and its potential therapeutic applications.