Levosimendan was purchased from Wako Pure Chemical Industries (Osaka, Japan). Levosimendan was resolved in dimethyl sulfoxide (DMSO) and stored at −80 °C. For GalN/LPS experiments, resolved levosimendan was diluted by 1 mL normal saline for each rat that the concentration of DMSO was decided at 2%. Recombinant human interleukin-1β (IL-1β; 2 × 107 U/mg protein) was purchased from MyBioSource (San Diego, CA, USA). Isoflurane, pentobarbital sodium, collagenase, a Transaminase CII-test kit, GalN, 10% formalin, and a PicaGene Luminescence kit were obtained from Wako Pure Chemical Industries (Osaka, Japan). LPS (Escherichia coli; O111:B4) and mouse anti-β-tubulin were obtained from Sigma–Aldrich (St. Louis, MO, USA). Enzyme-linked immunosorbent assay (ELISA) kits were obtained from Life Technologies (Carlsbad, CA, USA). TRIzol Reagent was obtained from Thermo Scientific (Waltham, MA, USA). T4 polynucleotide kinase, Oligo (dT) Primer, dNTPs Mixture, RNase Inhibitor, and Rever Tra Ace were obtained from Toyobo (Osaka, Japan). Beta-Glo kits and mouse immunoglobulin κ light chain were obtained from Promega (Madison, WI, USA).
Picagene luminescence kit
Manufactured by Fujifilm
Sourced in Japan
The PicaGene Luminescence kit is a laboratory equipment product designed for the detection and quantification of luminescent signals. The kit includes essential components for performing luminescence-based assays, enabling researchers to measure and analyze light-emitting reactions in their experiments.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Beta glo kit, by Promega (2 mentions)
Formalin, by Fujifilm (2 mentions)
T4 polynucleotide kinase, by Toyobo (2 mentions)
Transaminase cii test kit, by Fujifilm (2 mentions)
Rnase inhibitor, by Toyobo (2 mentions)
Revertra ace, by Toyobo (2 mentions)
Isoflurane, by Fujifilm (2 mentions)
Collagenase, by Fujifilm (2 mentions)
Pentobarbital sodium, by Fujifilm (2 mentions)
Mouse immunoglobulin κ light chain, by Promega (2 mentions)
Mouse anti β tubulin, by Merck Group (2 mentions)
Lps escherichia coli o111 b4, by Merck Group (2 mentions)
Dntps mixture, by Toyobo (2 mentions)
Enzyme linked immunosorbent assay elisa kit, by Thermo Fisher Scientific (1 mentions)
Oligo dt primer, by Toyobo (1 mentions)
X tremegene 9 dna transfection reagent, by Merck Group (1 mentions)
Phusion high fidelity dna polymerase, by New England Biolabs (1 mentions)
4 protocols using picagene luminescence kit
1
Levosimendan Inhibits GalN/LPS-induced Liver Injury
2
Hepatoprotective Effects of OMZ and IL-1β
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OMZ (20 mg) and recombinant human IL-1β (2 × 107 U/mg protein) were purchased from Nichi-iko Co, Ltd. (Toyama, Japan) and MyBioSource (San Diego, Calif). Isoflurane, pentobarbital sodium, collagenase, Transaminase CII-test kit, GalN, 10% formalin, and PicaGene Luminescence kit were from Wako Pure Chemical Industries (Osaka, Japan). LPS (Escherichia coli; O111:B4) and mouse anti-β-tubulin were from Sigma-Aldrich Japan (Tokyo, Japan). Enzyme-linked immunosorbent assay (kits were from Life Technologies Japan (Tokyo, Japan). TRIzol Reagent was from Thermo Scientific (Waltham, Mass). T4 polynucleotide kinase, Oligo (dT) Primer (25 ng), dNTPs Mixture, RNase Inhibitor, and Rever Tra Ace were from Toyobo (Osaka, Japan). Beta-Glo kits and mouse immunoglobulin κ light chain were from Promega (Fitchburg, Wis).
3
Luminescence-based Antiparasitic Screening
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Luc2-Tulahuen epimastigotes (2 × 105/well) were dispensed into 96-well plate. After 72 h incubation, luminescence intensity was measured by adding lysis buffer with luciferin as substrate (PicaGene Luminescence Kit, Fuji Film Wako chemicals, Japan) (Measurement time: 1 s) [24 (link)].
The IC50 was calculated using the following equation: where A is the lowest concentration at which the percentage inhibition exceeds 50%, B is the highest concentration at which percentage inhibition is less than 50%, and C and D are the percentage inhibition of the sample at concentrations B and A, respectively.
The IC50 was calculated using the following equation: where A is the lowest concentration at which the percentage inhibition exceeds 50%, B is the highest concentration at which percentage inhibition is less than 50%, and C and D are the percentage inhibition of the sample at concentrations B and A, respectively.
4
Promoter Activity Evaluation via Luciferase Assay
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To evaluate the activity of the promoter region, a luciferase reporter assay was performed as previously described29 (link). Briefly, the promoter region of interest (−190 to +41 from the cap site of the gene) was amplified by PCR using Phusion High-Fidelity DNA polymerase (New England BioLabs, Ipswich, MA, USA) and restriction site–tagged primers (NheI site upstream and HindIII site downstream). The PCR product was cloned between the NheI and HindIII sites of a luciferase reporter plasmid, pGL4.17 (Promega Corp., Fitchburg, WI, USA). The resulting plasmid was transfected into WERI Rb1 cells using X-tremeGENE 9 DNA transfection reagent (Sigma-Aldrich, St. Louis, MO, USA). Two days after transfection, the cells were collected and lysed using PicaGene Cell Culture Lysis Reagent Luc (Wako Chemicals, Osaka, Japan). Luciferase activity was measured using a luminometer (Lumicounter NU-2500, Microtech Co., Ltd., Funahashi, Japan) and PicaGene Luminescence kit (Wako). Transfection efficiency was monitored in cells co-transfected with a ß-galactosidase–encoding plasmid (Promega).
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