Nude mice established with subcutaneous U87ΔEGFR or orthotopic MDA468 tumors were treated with control IgG or OS2966 (5 mg/kg) by intraperitoneal injection at days −2, 0, 2, and 4. Two days post control IgG or OS2966 treatment, the mice were treated with rHSVQ1-Luc of 1 × 105 pfu by intratumoral injection and subjected to in vivo luciferase imaging at days 1, 2, 3, 4, and 5 after the oHSV injection. Briefly, mice were injected with Luciferin substrate solution (25 mg/ml in PBS, dose of 100 mg/kg, Perkin Elmer, Waltham, MA) by an intraperitoneal injection followed by anesthesia. The anesthetized mice were placed on non-fluorescent black paper on the imaging platform under ZFOV-24 zoom lens-installed IVIS Lumina Series III Pre-clinical In Vivo Imaging System (Perkin Elmer, Waltham, MA) to reduce background noise. Luminescence intensity was expressed as Averaged Radiance [p/s/cm2/sr], then normalized by tumor volume (mm3) obtained by MRI.
Luciferin substrate solution
Manufactured by PerkinElmer
Luciferin substrate solution is a laboratory reagent used in luminescence-based assays. It contains the luciferin compound, which is a substrate for the luciferase enzyme. The solution is typically used in applications that require the detection and quantification of luciferase activity, such as reporter gene assays and cell-based assays.
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2 protocols using luciferin substrate solution
1
Efficacy of OS2966 and oHSV Therapy
2
In Vitro and In Vivo Luciferase Assay for Gene Expression
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In vitro luciferase assay was performed by cell transfection. Briefly, Hela cells were plated at 1 × 105/well in 96well plate and then transfected with equimolar concentrations of DNA plasmid or DNA amplicon (1 and 0.3 μg). After 24 h, luciferase expression signal was measured by Imaging at an IVIS 200 imaging device (Perkin Elmer, USA).
In order to assess luciferase expression in vivo, female BALB/c mice (5 mice/group) were anesthetized using 97% oxygen and 3% isoflurane (Isoba, UK) then injected by EP in quadriceps muscle with a DNA plasmid encoding Luciferase (pcDNA3-Hygro-Luc at 1 μg/mouse) or equimolar concentrations of DNA amplicon. Imaging was performed under gas anesthesia at IVIS 200 imaging system at 24 h, 48 h and one week after injection, 8 min after injecting subcutaneously a luciferin substrate solution (15 mg/ml, Perkin Elmer) at 10μl/g of body weight.
In order to assess luciferase expression in vivo, female BALB/c mice (5 mice/group) were anesthetized using 97% oxygen and 3% isoflurane (Isoba, UK) then injected by EP in quadriceps muscle with a DNA plasmid encoding Luciferase (pcDNA3-Hygro-Luc at 1 μg/mouse) or equimolar concentrations of DNA amplicon. Imaging was performed under gas anesthesia at IVIS 200 imaging system at 24 h, 48 h and one week after injection, 8 min after injecting subcutaneously a luciferin substrate solution (15 mg/ml, Perkin Elmer) at 10μl/g of body weight.
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