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Vasopressin

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Vasopressin is a laboratory product that functions as a nonapeptide hormone. It is involved in the regulation of water absorption and blood pressure.

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Lab products found in correlation

3 isobutyl 1 methylxanthine ibmx, by Merck Group (2 mentions) Ptre2 hygromycin vector, by Thermo Fisher Scientific (2 mentions) Rx821002, by Bio-Techne (2 mentions) L phenyalanine, by Merck Group (2 mentions) Sb206553, by Bio-Techne (2 mentions) Gr127935, by Bio-Techne (2 mentions) Nsd1015, by Merck Group (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)

4 protocols using vasopressin

1

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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2

Detailed ACSF compositions for in vitro experiments

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Two kinds of ACSF were used in in vitro experiments: mACSF in the dissection dish before recording and nACSF in the recording chamber. mACSF was composed of (in mM) 118 NaCl, 24 NaHCO3, 1.5 CaCl2, 3 KCl, 5 MgCl2, 1.4 NaH2PO4, 1.3 MgSO4, 25 d-glucose, and 1 kynurenic acid. nACSF was composed of (in mM) 122 NaCl, 25 NaHCO3, 2.5 CaCl2, 3 KCl, 1 MgCl2, 0.5 NaH2PO4, and 12 d-glucose. Both types of ACSF were saturated with carbogen (95% O2-5% CO2) and maintained at pH 7.4. The drugs added to the nACSF were 5-HTP, clorgyline, pargyline, tryptamine, L-tryptophan, tyramine, L-tyrosine, 2-phenylethylamine, L-phenyalanine, NSD1015 (Sigma-Aldrich), α-methyl-5-HT, octopamine, SB206553, RX821002, GR127935, dobutamine, vasopressin (Tocris) and zolmitriptan (AstraZenica). All drugs were first dissolved as a 10 – 50 mM stock solution in water before final dilution in ACSF for in vitro ventral root reflexes recording and DIC microscopy or in saline for in vivo oxygen measurements and two photon microscopy, with the exception of zolmitriptan, which was dissolved in minimal amounts of DMSO (DMSO final concentration in nACSF was 0.04%). DMSO alone had no effect on in vitro LLR in vehicle controls, as compare to nACSF control state.
Li Y., Lucas-Osma A.M., Black S., Bandet M.V., Stephens M.J., Vavrek R., Sanelli L., Fenrich K.K., Di Narzo A.F., Dracheva S., Winship I.R., Fouad K, & Bennett D.J. (2017). Pericytes impair capillary blood flow and motor function after chronic spinal cord injury. Nature medicine, 23(6), 733-741.
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3

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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4

Detailed ACSF compositions for in vitro experiments

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Two kinds of ACSF were used in in vitro experiments: mACSF in the dissection dish before recording and nACSF in the recording chamber. mACSF was composed of (in mM) 118 NaCl, 24 NaHCO3, 1.5 CaCl2, 3 KCl, 5 MgCl2, 1.4 NaH2PO4, 1.3 MgSO4, 25 d-glucose, and 1 kynurenic acid. nACSF was composed of (in mM) 122 NaCl, 25 NaHCO3, 2.5 CaCl2, 3 KCl, 1 MgCl2, 0.5 NaH2PO4, and 12 d-glucose. Both types of ACSF were saturated with carbogen (95% O2-5% CO2) and maintained at pH 7.4. The drugs added to the nACSF were 5-HTP, clorgyline, pargyline, tryptamine, L-tryptophan, tyramine, L-tyrosine, 2-phenylethylamine, L-phenyalanine, NSD1015 (Sigma-Aldrich), α-methyl-5-HT, octopamine, SB206553, RX821002, GR127935, dobutamine, vasopressin (Tocris) and zolmitriptan (AstraZenica). All drugs were first dissolved as a 10 – 50 mM stock solution in water before final dilution in ACSF for in vitro ventral root reflexes recording and DIC microscopy or in saline for in vivo oxygen measurements and two photon microscopy, with the exception of zolmitriptan, which was dissolved in minimal amounts of DMSO (DMSO final concentration in nACSF was 0.04%). DMSO alone had no effect on in vitro LLR in vehicle controls, as compare to nACSF control state.
Li Y., Lucas-Osma A.M., Black S., Bandet M.V., Stephens M.J., Vavrek R., Sanelli L., Fenrich K.K., Di Narzo A.F., Dracheva S., Winship I.R., Fouad K, & Bennett D.J. (2017). Pericytes impair capillary blood flow and motor function after chronic spinal cord injury. Nature medicine, 23(6), 733-741.
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