Activation of phospholipase C was determined by quantification of the [3H]-inositol phosphates (InsP) [21 (link)]. NFκB-luciferase reporter gene assays were performed in HEK293T cells [52 (link)] and luminescence (1 s per well) was measured with a Mithras LB940 multilabel plate reader (Berthold Technologies). VEGF levels were measured in conditioned medium by ELISA using the Quantikine human-VEGF ELISA kit (R&D systems) [25 (link)]. In all VEGF ELISA assays, the lower detection limit was 15 pg/ml.
Mithras lb940 multilabel plate reader
Manufactured by Berthold Technologies
Sourced in United States
The Mithras LB940 is a multilabel plate reader designed for a variety of applications. It is capable of performing measurements such as absorbance, fluorescence, and luminescence in microplates.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using mithras lb940 multilabel plate reader
1
Quantification of Phospholipase C Activation and NFκB Signaling
2
Assessing GPCR-Arrestin Interactions
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HEK293T cells were transfected
with 0.5
μg of WT or mutant H4R-Rluc8 in combination with
4 μg of β-arrestin1-eYFP, β-arrestin2-mVenus, or
GRK2/3/5/6-mVenus DNA, or 2 μg of Venus-Rab5a DNA. At 24 h post-transfection,
cells were transferred to poly-l -lysine-coated white 96-well
plates (Greiner Bio-one; Frickenhausen, Germany). At 48 h post-transfection,
baseline BRET was measured using 5 μM coelenterazine-H (Promega;
Madison, WI, USA) in HBSS in a Mithras LB940 multilabel plate reader
(Berthold Technologies; Bad Wildbad, Germany) followed by stimulation
with H4R ligands at 37 °C, as previously described.67 (link) The BRET ratio was calculated by dividing acceptor
light emission at 540 nm by Rluc8 light emission at 480 nm.
with 0.5
μg of WT or mutant H4R-Rluc8 in combination with
4 μg of β-arrestin1-eYFP, β-arrestin2-mVenus, or
GRK2/3/5/6-mVenus DNA, or 2 μg of Venus-Rab5a DNA. At 24 h post-transfection,
cells were transferred to poly-
plates (Greiner Bio-one; Frickenhausen, Germany). At 48 h post-transfection,
baseline BRET was measured using 5 μM coelenterazine-H (Promega;
Madison, WI, USA) in HBSS in a Mithras LB940 multilabel plate reader
(Berthold Technologies; Bad Wildbad, Germany) followed by stimulation
with H4R ligands at 37 °C, as previously described.67 (link) The BRET ratio was calculated by dividing acceptor
light emission at 540 nm by Rluc8 light emission at 480 nm.
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