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Promega glosensor 22f plasmid
Glosensor 22f Plasmid, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/glosensor+22f+plasmid/pglosensor+22f+camp+plasmid/pm40537032-161-16-19
Average 90 stars, based on 1 article reviews
glosensor 22f plasmid - by Bioz Stars, 2026-09
90/100 stars

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cAMP Assay:

Article Title: MC4R Variants Modulate α-MSH and Setmelanotide Induced Cellular Signaling at Multiple Levels
Article Snippet: The pcDNA3.1(+)-MC4R expression plasmid, containing the WT human MC4R cDNA was obtained from the cDNA Resource Center ( www.cdna.org , USA). .. For the GloSensor cAMP assay, pcDNA3.1(+) expression plasmid containing WT or variant MC4R and the GloSensor 22F plasmid (Promega, USA) were used to measure cAMP accumulation. .. For the β-arrestin-2 recruitment assay, the cDNA of human ARRB2 (β-arrestin-2; SinoBiological Europe GmbH, Germany) was cloned into the pBiT2.1-N [TK/SmBiT] vector, generating an N-terminally tagged ARRB2, and the cDNA of human MC4R was cloned into pBiT1.1-C [TK/LgBiT] vector (Promega, USA), generating a C-terminally LgBiT-tagged MC4R.

Expressing:

Article Title: MC4R Variants Modulate α-MSH and Setmelanotide Induced Cellular Signaling at Multiple Levels
Article Snippet: The pcDNA3.1(+)-MC4R expression plasmid, containing the WT human MC4R cDNA was obtained from the cDNA Resource Center ( www.cdna.org , USA). .. For the GloSensor cAMP assay, pcDNA3.1(+) expression plasmid containing WT or variant MC4R and the GloSensor 22F plasmid (Promega, USA) were used to measure cAMP accumulation. .. For the β-arrestin-2 recruitment assay, the cDNA of human ARRB2 (β-arrestin-2; SinoBiological Europe GmbH, Germany) was cloned into the pBiT2.1-N [TK/SmBiT] vector, generating an N-terminally tagged ARRB2, and the cDNA of human MC4R was cloned into pBiT1.1-C [TK/LgBiT] vector (Promega, USA), generating a C-terminally LgBiT-tagged MC4R.

Article Title: Mammalian type opsin 5 preferentially activates G14 in Gq-type G proteins triggering intracellular calcium response
Article Snippet: 293T cells were seeded into white-walled 96-well plates (MS8096W, Sumitomo Bakelite) at 20,000 cells per well in DMEM/F-12 (FUJIFILM Wako) containing 10% FBS. .. After a day, cells were transfected with 0.05 μg cOpn5L2 expression plasmid and 0.05 μg Glosensor 22F plasmid (Promega) using 0.4 μg polyethyleneimine (PEI) (PEI MAX, Polyscience) diluted in OptiMEM (Thermo Scientific). ..

Plasmid Preparation:

Article Title: MC4R Variants Modulate α-MSH and Setmelanotide Induced Cellular Signaling at Multiple Levels
Article Snippet: The pcDNA3.1(+)-MC4R expression plasmid, containing the WT human MC4R cDNA was obtained from the cDNA Resource Center ( www.cdna.org , USA). .. For the GloSensor cAMP assay, pcDNA3.1(+) expression plasmid containing WT or variant MC4R and the GloSensor 22F plasmid (Promega, USA) were used to measure cAMP accumulation. .. For the β-arrestin-2 recruitment assay, the cDNA of human ARRB2 (β-arrestin-2; SinoBiological Europe GmbH, Germany) was cloned into the pBiT2.1-N [TK/SmBiT] vector, generating an N-terminally tagged ARRB2, and the cDNA of human MC4R was cloned into pBiT1.1-C [TK/LgBiT] vector (Promega, USA), generating a C-terminally LgBiT-tagged MC4R.

Article Title: Real-Time Signaling Assays Demonstrate Somatostatin Agonist Bias for Ion Channel Regulation in Somatotroph Tumor Cells
Article Snippet: .. Reagents were purchased from the following sources: DMEM and F12 medium from Lonza (Allendale, NJ); CO 2 -independent media from Gibco/ThermoFisher (Waltham, MA); fetal bovine serum (FBS) and horse serum from Atlanta Biologicals (Flowery Branch, GA); pertussis toxin (PTX) from List Biological Laboratories Inc. (Campbell, CA); the water soluble forskolin analogue NKH477 from Cayman Chemical Company (Ann Arbor, MI); GloSensor 22F plasmid and FuGENE transfection reagent from Promega (Madison, WI); and D-Luciferin from GoldBio (St. Louis, MO). .. Anti-HA epitope antibody was purchased from Covance (Princeton, NJ), and goat anti-mouse secondary antibody was ordered from KPL (Gaithersburg, MD).

Article Title: Mammalian type opsin 5 preferentially activates G14 in Gq-type G proteins triggering intracellular calcium response
Article Snippet: 293T cells were seeded into white-walled 96-well plates (MS8096W, Sumitomo Bakelite) at 20,000 cells per well in DMEM/F-12 (FUJIFILM Wako) containing 10% FBS. .. After a day, cells were transfected with 0.05 μg cOpn5L2 expression plasmid and 0.05 μg Glosensor 22F plasmid (Promega) using 0.4 μg polyethyleneimine (PEI) (PEI MAX, Polyscience) diluted in OptiMEM (Thermo Scientific). ..

Article Title: Estimation of Dopamine D 1 Receptor Agonist Binding Kinetics Using Time-Resolved Functional Assays: Relation to Agonist-Induced Receptor Internalization by Investigational Antiparkinsonian Therapeutics.
Article Snippet: .. HEK293T cells were grown in 10 cm dishes and transfected as described above with 6 μg GloSensor 22F plasmid (Promega, Madison, WI) and 1 μg D1R plasmid in pXOOM, together with empty pcDNA3.1 as needed to reach 20 μg plasmid DNA per dish. .. The cAMP assay was performed essentially as previously described.32,57 In brief, 24 h after transfection, cells were lifted off, pelleted, and resuspended counted and further diluted to 500,000 cells ml−1 in HBSS supplemented with 300 μM 3-isobutyl-1methylxanthine (IBMX; Sigma-Aldrich) and GloSensor reagent (Promega) and 1 mM ascorbic acid.

Article Title: In Vitro Comparison of Ulotaront (SEP-363856) and Ralmitaront (RO6889450): Two TAAR1 Agonist Candidate Antipsychotics.
Article Snippet: On the day of the assay, cells were resuspended in dPBS with 10 μM h-Coelenterazine (NanoLight Technologies) and seeded in a volume of 90 μL per well into white 96-well plates (Corning). . Luminescence signal counts were normalized as fold change relative to vehicle-treated control wells. cAMP Assay HEK293T cells were grown in 5% CO2 atmosphere at 37°C on 10-cm dishes (VWR part of Avantor, Radnor, PA, USA) containing Dulbecco’s modified eagle medium with GlutaMAX (ThermoFischer Scientific) supplemented with 0.01% penicillin-streptomycin (ThermoFisher Scientific) and 10% fetal bovine serum (ThermoFisher Scientific). .. Cells were transfected using linear polyethylenimine (Polysciences Inc., Warrington, PA, USA) with 6 μg GloSensor 22F plasmid (Promega, Madison, WI, USA) and 4 μg (TAAR1) or 1 μg (5-HT1AR and D2R) of receptor together with empty pcDNA3.1 as needed to reach 20 μg plasmid DNA per dish. .. Twenty-four hours after transfection, cells were lifted off, pelleted, and resuspended in Hank’s buffered salt solution supplemented with 300 μM 3-isobutyl-1-methylxanthine (SigmaAldrich) and GloSensor reagent (Promega).

Article Title: The ubiquitination status of the glucagon receptor determines signal bias.
Article Snippet: The quantification of protein bands was done by using Image Lab software (Bio-Rad; https://www.bio-rad.com/en-us/product/ image-lab-software?ID=KRE6P5E8Z). .. GloSensor assay for determining cAMP production HEK-293 cells transiently transfected either with GCGR-WT, GCGR single lysine mutants or GCGR-5KR and GloSensor 22F plasmid (Promega) in 6-well dishes. ..

Article Title: In Vitro Comparison of Ulotaront (SEP-363856) and Ralmitaront (RO6889450): Two TAAR1 Agonist Candidate Antipsychotics
Article Snippet: HEK293T cells were grown in 5% CO 2 atmosphere at 37°C on 10-cm dishes (VWR part of Avantor, Radnor, PA, USA) containing Dulbecco’s modified eagle medium with GlutaMAX (ThermoFischer Scientific) supplemented with 0.01% penicillin-streptomycin (ThermoFisher Scientific) and 10% fetal bovine serum (ThermoFisher Scientific). .. Cells were transfected using linear polyethylenimine (Polysciences Inc., Warrington, PA, USA) with 6 μg GloSensor 22F plasmid (Promega, Madison, WI, USA) and 4 μg (TAAR1) or 1 μg (5-HT 1A R and D 2 R) of receptor together with empty pcDNA3.1 as needed to reach 20 μg plasmid DNA per dish. .. Twenty-four hours after transfection, cells were lifted off, pelleted, and resuspended in Hank’s buffered salt solution supplemented with 300 μM 3-isobutyl-1-methylxanthine (Sigma-Aldrich) and GloSensor reagent (Promega).

Article Title: The ubiquitination status of the glucagon receptor determines signal bias
Article Snippet: The quantification of protein bands was done by using Image Lab software (Bio-Rad; https://www.bio-rad.com/en-us/product/image-lab-software?ID=KRE6P5E8Z ). .. HEK-293 cells transiently transfected either with GCGR-WT, GCGR single lysine mutants or GCGR-5KR and GloSensor 22F plasmid (Promega) in 6-well dishes. ..

Variant Assay:

Article Title: MC4R Variants Modulate α-MSH and Setmelanotide Induced Cellular Signaling at Multiple Levels
Article Snippet: The pcDNA3.1(+)-MC4R expression plasmid, containing the WT human MC4R cDNA was obtained from the cDNA Resource Center ( www.cdna.org , USA). .. For the GloSensor cAMP assay, pcDNA3.1(+) expression plasmid containing WT or variant MC4R and the GloSensor 22F plasmid (Promega, USA) were used to measure cAMP accumulation. .. For the β-arrestin-2 recruitment assay, the cDNA of human ARRB2 (β-arrestin-2; SinoBiological Europe GmbH, Germany) was cloned into the pBiT2.1-N [TK/SmBiT] vector, generating an N-terminally tagged ARRB2, and the cDNA of human MC4R was cloned into pBiT1.1-C [TK/LgBiT] vector (Promega, USA), generating a C-terminally LgBiT-tagged MC4R.

Transfection:

Article Title: Real-Time Signaling Assays Demonstrate Somatostatin Agonist Bias for Ion Channel Regulation in Somatotroph Tumor Cells
Article Snippet: .. Reagents were purchased from the following sources: DMEM and F12 medium from Lonza (Allendale, NJ); CO 2 -independent media from Gibco/ThermoFisher (Waltham, MA); fetal bovine serum (FBS) and horse serum from Atlanta Biologicals (Flowery Branch, GA); pertussis toxin (PTX) from List Biological Laboratories Inc. (Campbell, CA); the water soluble forskolin analogue NKH477 from Cayman Chemical Company (Ann Arbor, MI); GloSensor 22F plasmid and FuGENE transfection reagent from Promega (Madison, WI); and D-Luciferin from GoldBio (St. Louis, MO). .. Anti-HA epitope antibody was purchased from Covance (Princeton, NJ), and goat anti-mouse secondary antibody was ordered from KPL (Gaithersburg, MD).

Article Title: Mammalian type opsin 5 preferentially activates G14 in Gq-type G proteins triggering intracellular calcium response
Article Snippet: 293T cells were seeded into white-walled 96-well plates (MS8096W, Sumitomo Bakelite) at 20,000 cells per well in DMEM/F-12 (FUJIFILM Wako) containing 10% FBS. .. After a day, cells were transfected with 0.05 μg cOpn5L2 expression plasmid and 0.05 μg Glosensor 22F plasmid (Promega) using 0.4 μg polyethyleneimine (PEI) (PEI MAX, Polyscience) diluted in OptiMEM (Thermo Scientific). ..

Article Title: Estimation of Dopamine D 1 Receptor Agonist Binding Kinetics Using Time-Resolved Functional Assays: Relation to Agonist-Induced Receptor Internalization by Investigational Antiparkinsonian Therapeutics.
Article Snippet: .. HEK293T cells were grown in 10 cm dishes and transfected as described above with 6 μg GloSensor 22F plasmid (Promega, Madison, WI) and 1 μg D1R plasmid in pXOOM, together with empty pcDNA3.1 as needed to reach 20 μg plasmid DNA per dish. .. The cAMP assay was performed essentially as previously described.32,57 In brief, 24 h after transfection, cells were lifted off, pelleted, and resuspended counted and further diluted to 500,000 cells ml−1 in HBSS supplemented with 300 μM 3-isobutyl-1methylxanthine (IBMX; Sigma-Aldrich) and GloSensor reagent (Promega) and 1 mM ascorbic acid.

Article Title: In Vitro Comparison of Ulotaront (SEP-363856) and Ralmitaront (RO6889450): Two TAAR1 Agonist Candidate Antipsychotics.
Article Snippet: On the day of the assay, cells were resuspended in dPBS with 10 μM h-Coelenterazine (NanoLight Technologies) and seeded in a volume of 90 μL per well into white 96-well plates (Corning). . Luminescence signal counts were normalized as fold change relative to vehicle-treated control wells. cAMP Assay HEK293T cells were grown in 5% CO2 atmosphere at 37°C on 10-cm dishes (VWR part of Avantor, Radnor, PA, USA) containing Dulbecco’s modified eagle medium with GlutaMAX (ThermoFischer Scientific) supplemented with 0.01% penicillin-streptomycin (ThermoFisher Scientific) and 10% fetal bovine serum (ThermoFisher Scientific). .. Cells were transfected using linear polyethylenimine (Polysciences Inc., Warrington, PA, USA) with 6 μg GloSensor 22F plasmid (Promega, Madison, WI, USA) and 4 μg (TAAR1) or 1 μg (5-HT1AR and D2R) of receptor together with empty pcDNA3.1 as needed to reach 20 μg plasmid DNA per dish. .. Twenty-four hours after transfection, cells were lifted off, pelleted, and resuspended in Hank’s buffered salt solution supplemented with 300 μM 3-isobutyl-1-methylxanthine (SigmaAldrich) and GloSensor reagent (Promega).

Article Title: The ubiquitination status of the glucagon receptor determines signal bias.
Article Snippet: The quantification of protein bands was done by using Image Lab software (Bio-Rad; https://www.bio-rad.com/en-us/product/ image-lab-software?ID=KRE6P5E8Z). .. GloSensor assay for determining cAMP production HEK-293 cells transiently transfected either with GCGR-WT, GCGR single lysine mutants or GCGR-5KR and GloSensor 22F plasmid (Promega) in 6-well dishes. ..

Article Title: In Vitro Comparison of Ulotaront (SEP-363856) and Ralmitaront (RO6889450): Two TAAR1 Agonist Candidate Antipsychotics
Article Snippet: HEK293T cells were grown in 5% CO 2 atmosphere at 37°C on 10-cm dishes (VWR part of Avantor, Radnor, PA, USA) containing Dulbecco’s modified eagle medium with GlutaMAX (ThermoFischer Scientific) supplemented with 0.01% penicillin-streptomycin (ThermoFisher Scientific) and 10% fetal bovine serum (ThermoFisher Scientific). .. Cells were transfected using linear polyethylenimine (Polysciences Inc., Warrington, PA, USA) with 6 μg GloSensor 22F plasmid (Promega, Madison, WI, USA) and 4 μg (TAAR1) or 1 μg (5-HT 1A R and D 2 R) of receptor together with empty pcDNA3.1 as needed to reach 20 μg plasmid DNA per dish. .. Twenty-four hours after transfection, cells were lifted off, pelleted, and resuspended in Hank’s buffered salt solution supplemented with 300 μM 3-isobutyl-1-methylxanthine (Sigma-Aldrich) and GloSensor reagent (Promega).

Article Title: The ubiquitination status of the glucagon receptor determines signal bias
Article Snippet: The quantification of protein bands was done by using Image Lab software (Bio-Rad; https://www.bio-rad.com/en-us/product/image-lab-software?ID=KRE6P5E8Z ). .. HEK-293 cells transiently transfected either with GCGR-WT, GCGR single lysine mutants or GCGR-5KR and GloSensor 22F plasmid (Promega) in 6-well dishes. ..



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KEY RESOURCES TABLE
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KEY RESOURCES TABLE

Journal: Neuron

Article Title: In vivo photopharmacology with light-activated opioid drugs

doi: 10.1016/j.neuron.2023.09.017

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Media in 10 cm TC dishes with HEK 293T cells (at around 70% confluence) were replaced with Opti-MEM (Invitrogen) followed by addition of a mixture of the GPCR plasmid, the GloSensor (Promega) 22F cAMP dependent reporter plasmid (Promega), and Lipofectamine 2000 (Invitrogen) in Opti-MEM.

Techniques: Virus, Recombinant, cAMP Assay, Software