Ptre2 hygromycin vector

Manufactured by Thermo Fisher Scientific

The PTRE2-Hygromycin vector is a plasmid DNA construct designed for gene expression studies in mammalian cell lines. It contains a hygromycin resistance gene, allowing for selection of cells successfully transfected with the vector.

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Lab products found in correlation

3 isobutyl 1 methylxanthine ibmx, by Merck Group (2 mentions) Vasopressin, by Bio-Techne (2 mentions) Vasopressin, by PerkinElmer (1 mentions) Net 114a, by PerkinElmer (1 mentions) Pierce pre coated iodination tubes, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Hygromycin (503 citations)

2 protocols using ptre2 hygromycin vector

Characterization of V2R Pharmacoperones

Cited 2 times

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SR121463B is a V2R antagonist and known pharmacoperone that was used in the current study, after being generously provided by Dr. Claudine Serradeil at Sanofi-Aventis and used as received. Other pharmacoperones were identified by us by high throughput screening of a large chemical library [22 (link), 23 (link)]. Several reagents were used as obtained from indicated vendors: 3-Isobutyl-1-methylxanthine (IBMX, Sigma Aldrich, St. Louis, MO), vasopressin (Tocris Biosciences, Bristol, England UK), fetal calf serum (FCS, Hyclone, Logan, UT), Dulbecco’s MEM (DMEM), PBS (GIBCO, Invitrogen). pTRE2-Hygromycin vector (Invitrogen, San Diego, CA), human arginine-vasopressin 2 receptor (V2R; Gene Bank Accession Number: AY242131), Gq plasmid (Gene Bank Accession Number: U43083) [24 (link)]; both plasmids from cDNA Resource Center; www.cdna.org;), myo-[2-³H(N)]-inositol (NET-114A; PerkinElmer, Waltham, MA), vasopressin (8-L-arginine), [phenylalaninyl-3,4,5-³H(N)- (NET800, specific activity = 66.3 Ci/mmol; PerkinElmer, Waltham, MA), and unbound 125-Iodine (016303710; MP Biomedicals, Santa Ana, CA).

Janovick J.A., Spicer T.P., Bannister T.D., Scampavia L, & Conn P.M. (2017). Pharmacoperone rescue of vasopressin 2 receptor mutants reveals unexpected constitutive activity and coupling bias. PLoS ONE, 12(8), e0181830.

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Pharmacoperone Antagonist Assay Protocol

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SR121463B, a V2R peptidomimetic antagonist used in the current study as a known pharmacoperone drug, was generously provided by Dr. Claudine Serradeil at Sanofi-Aventis and used as received. Test compounds used in this study were prepared at the screening facility and stored as 10 mM DMSO stock solutions at −20 °C in sealed polypropylene plates and these stocks were also used for the orthologous assay. 3-Isobutyl-1-methylxanthine (IBMX, Sigma Aldrich, St. Louis, MO), vasopressin (Tocris Biosciences, Bristol, England UK) and fetal bovine serum (FBS, Hyclone, Logan, UT) were obtained as indicated. The V2 receptor antagonist, d(CH₂)5[D-Ile(2),Ile(4),Tyr-NH₂(9)]AVP was a kind gift of Maurice Manning (9 (link), 10 (link)) and was radiolabeled using Pierce Pre-Coated Iodination Tubes (Thermo Fisher Scientific, Waltham, MA) and 125-Iodine (NEZ033L; PerkinElmer, Waltham, MA), DMEM, PBS (GIBCO, Invitrogen). pTRE2-Hygromycin vector (Invitrogen, San Diego, CA), myo-[2-³H(N)]-inositol (NET-114A; PerkinElmer, Waltham, MA) were obtained as indicated.

Janovick J.A., Spicer T.P., Smith E., Bannister T.D., Kenakin T., Scampavia L, & Conn P.M. (2016). Receptor Antagonism/Agonism Can Be Uncoupled from Pharmacoperone Activity. Molecular and cellular endocrinology, 434, 176-185.

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