Htrf ip1 kit

Manufactured by PerkinElmer

The HTRF-IP1 kit is a fluorescence-based assay designed for the detection and quantification of inositol 1-phosphate (IP1) levels in cells. The kit utilizes the Homogeneous Time-Resolved Fluorescence (HTRF) technology to measure IP1 concentrations, which are an indirect indicator of G-protein coupled receptor (GPCR) activation. The kit provides a simple, sensitive, and high-throughput method for the investigation of GPCR signaling pathways.

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Lab products found in correlation

Mithras lb940 reader, by Berthold Technologies (1 mentions) Htrf camp dynamic kit, by PerkinElmer (1 mentions)

Spelling variants of this product

HTRF kit (16 citations) HTRF-IP1 assay kit (4 citations)

2 protocols using htrf ip1 kit

Quantifying cAMP and IP1 Dynamics

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Changes of the intracellular second messengers cAMP and IP1 were quantified with the HTRF-cAMP dynamic kit and the HTRF-IP1 kit, respectively (CisBio International), on a Mithras LB 940 reader (Berthold Technologies) according to the manufacturer’s instructions and as described elsewhere in detail (Schröder et al., 2009 (link); Schmidt et al., 2011 (link)). If BIM or its solvent were present during the assay, it was preincubated for 2 hr at 37°C.

Schmitz A.L., Schrage R., Gaffal E., Charpentier T.H., Wiest J., Hiltensperger G., Morschel J., Hennen S., Häußler D., Horn V., Wenzel D., Grundmann M., Büllesbach K.M., Schröder R., Brewitz H.H., Schmidt J., Gomeza J., Galés C., Fleischmann B.K., Tüting T., Imhof D., Tietze D., Gütschow M., Holzgrabe U., Sondek J., Harden T.K., Mohr K, & Kostenis E. (2014). A Cell-Permeable Inhibitor to Trap Gαq Proteins in the Empty Pocket Conformation. Chemistry & biology, 21(7), 890-902.

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Quantification of Intracellular IP1 Levels

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Intracellular alteration of the second messenger IP1 was quantified with the HTRF-IP1 kit (Cisbio Bioassays) following the manufacturer's instructions. Briefly, brown preadipocytes non-transduced and transduced with lentiviruses carrying GqQL (LVGqQL) or GFP (LVGFP) were cultivated for 48 h in growth medium (37 °C, 5% CO₂). For the assay, cells were resuspended in stimulation buffer containing 50 mM LiCl and transferred to a 384-well microtiter plate at a density of 12,500 cells per well. After 90 min incubation time, 3 μl of IP1-d2 conjugate followed by 3 μl of europium cryptate-labelled anti-IP1antibody solved in lysis buffer were added to the cells to quantify intracellular IP levels. After a further incubation of 1 h at room temperature, time-resolved fluorescence was measured at 620 and 665 nm with the Mithras LB 940 multimode reader.

Klepac K., Kilić A., Gnad T., Brown L.M., Herrmann B., Wilderman A., Balkow A., Glöde A., Simon K., Lidell M.E., Betz M.J., Enerbäck S., Wess J., Freichel M., Blüher M., König G., Kostenis E., Insel P.A, & Pfeifer A. (2016). The Gq signalling pathway inhibits brown and beige adipose tissue. Nature Communications, 7, 10895.

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