Collagenase I and Collagenase IV were purchased from Worthington Biochemical Corporation (Lakewood, NJ, USA). The mouse monoclonal anti-Tubulin was acquired at Sigma Aldrich (Milan, Italy), while the rabbit polyclonal anti-mGluR5 at EMD Millipore (Burlington, MA, USA). IP-One ELISA was provided by Cisbio (Codolet, France). Male Wistar rats were from Charles River Laboratories (Lecco, Italy).All other reagents were purchased from Sigma Aldrich (Milan, Italy). 1-Amino-1,3-dicarboxycyclopentane (ACPD) and 2-Methyl-6-(phenylethynyl)pyridine (MPEP) were purchased from Tocris Bioscience (Bristol, UK).
Ip one elisa
Manufactured by PerkinElmer
Sourced in France
The IP-One ELISA is a laboratory equipment product that measures the levels of inositol phosphate (IP) in cellular samples. It provides a quantitative assessment of IP production, which is an important indicator of cellular signaling activity.
Automatically generated - may contain errors
Lab products found in correlation
2 methyl 6 phenylethynyl pyridine mpep, by Bio-Techne (1 mentions)
Anti tubulin, by Merck Group (1 mentions)
Collagenase 4, by Worthington (1 mentions)
Collagenase 1, by Worthington (1 mentions)
Male wistar rats, by Charles River Laboratories (1 mentions)
Cyclic amp competitive elisa kit, by Thermo Fisher Scientific (1 mentions)
5 protocols using ip one elisa
1
Isolation and Analysis of mGluR5 in Rat
2
Quantifying PLC Signaling in Pancreatic Cells
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The activation of PLC signaling and inositol triphosphate (IP3) generation in Min6 cells and isolated islets was measured using the IP-One ELISA (CisBio), which quantifies the accumulation of inositol monophosphate (IP1), a downstream metabolite of IP3. Min6 cells, 15 mouse islets or 125 islet equivalent human islets, were incubated in stimulation buffer containing lithium chloride, supplemented with 16.7 mmol/L glucose, for 1 h at 37°C, in 5% CO2.
3
Measuring Ap-GnRH Signaling Pathways
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The accumulation of inositol 1-phosphate (IP1) was measured by the IP-One ELISA (Cisbio, Bedford, MA) according to the manufacturer’s instructions. S2 cells transfected as before with pAWG, pAWG-L or pAWG-S were treated without or with 10−12 M to 10−6 M of Ap-GnRH in a stimulation buffer provided by the kit for 3 h at 28°C. All treatments were performed in triplicates in 3 independent experiments. Proctolin (RYLPT; GenScript) was used at 10−6 M as a positive control for the assay. Proctolin receptor is endogenously expressed in S2 cells and its activation coupled to the activation of the IP pathway [22 (link)].
The accumulation of cAMP was measured by the Cyclic AMP Competitive ELISA Kit (ThermoFisher Scientific, Waltham, MA) using the acetylated protocol according to the manufacturer’s instructions. Transfected S2 cells were treated without or with 10−12 M to 10−6 M of Ap-GnRH for 1 hr at 28°C. All treatments were performed in triplicates in 3 independent experiments. Forskolin (5 μM) was used to show maximum stimulation, and 3-Isobutyl-1-methylxanthine (1 mM) was used to prevent cAMP degradation.
The accumulation of cAMP was measured by the Cyclic AMP Competitive ELISA Kit (ThermoFisher Scientific, Waltham, MA) using the acetylated protocol according to the manufacturer’s instructions. Transfected S2 cells were treated without or with 10−12 M to 10−6 M of Ap-GnRH for 1 hr at 28°C. All treatments were performed in triplicates in 3 independent experiments. Forskolin (5 μM) was used to show maximum stimulation, and 3-Isobutyl-1-methylxanthine (1 mM) was used to prevent cAMP degradation.
4
Measuring Bradykinin-Induced InsP1 in HASM Cells
5
Measuring Bradykinin-Induced InsP1 in HASM Cells
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HASM cells were seeded in 96-well plates and starved in serum-free media for 48 hours followed by 24 hours pre-treatment with vehicle or Af (2 µg ml−1). Cells were then stimulated with increasing concentrations of bradykinin for two minutes. InsP1 concentrations were measured by ELISA as described by the manufacturer (IP-One ELISA, Cisbio Bioassays).
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