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Rediject d luciferin ultra bioluminescent substrate

Manufactured by PerkinElmer

RediJect D-Luciferin Ultra Bioluminescent Substrate is a laboratory product that provides a luciferin substrate for bioluminescent applications. It is designed to enable the detection and quantification of luciferase-based reporter systems.

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6 protocols using rediject d luciferin ultra bioluminescent substrate

1

In Vivo Bioluminescent Imaging of Mice

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Prior to imaging, the mice were i.p. injected with 100 μL of 30 mg/ml RediJect D-Luciferin Ultra bioluminescent substrate (PerkinElmer). Mice were then anesthetized with isoflurane and imaged within 20 minutes for bioluminescence using the in vivo imager. All the bioluminescence data, except for those shown in Supplementary Figures 7d and e, were collected using the IVIS Spectrum In Vivo Imaging System (PerkinElmer), and bioluminescent photon outputs were quantified using the Living Image Software (PerkinElmer). The bioluminescence data shown in Supplementary Figures 7d and e were collected using the Lago X optical imaging system (Spectral Instruments Imaging) and analyzed using Aura software (Spectral Instruments Imaging).
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2

Establishing mir-155 Lymphoma Xenografts

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To establish mir-155LSLtTA lymphoma subcutaneous flank tumors, first enlarged spleens were extracted from 2–3 month-old mir-155LSLtTA mice with obvious lymphadenopathy (which generally correlated with incidence of splenomegaly). Using a 100 μm pore size cell strainer technique, spleen tissue was dispersed into a single cell suspension in 5% FBS in PBS on ice. Red blood cells were lysed using ammonium chloride lysis buffer (Stem Cell Technologies), and 5 × 106 cells were subcutaneously injected into nude mice. Tumors were generally palpable within 10 days; tumor volume was calculated as (Length x Width2)/2.
For bioluminescent xenograft tumors, KB cells were stably transfected with firefly luciferase and clonally selected via hygromycin B selection; 5 × 106 cells were subcutaneously injected into nude mice to establish tumors. RediJect D-Luciferin Ultra Bioluminescent Substrate (PerkinElmer) was administered via the manufacturer’s protocol for intravital monitoring of tumor bioluminescence using IVIS Spectrum (Caliper). It was pre-established that for all flank tumor studies, animals were excluded if their tumors had not reached a volume of 50–100 cm3 by the time of treatment. Animals were randomized into experimental arms by minimizing the differences in mean tumor size and standard deviation.
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3

Tongue Tumor Xenograft Model

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Nude mice (8 weeks of age) or WT and TNCKO (C57Bl/6J) mice (bred in house) were grafted in the first third part of the tongue. In particular, 3 × 106 OSCC13 cells in 10 µl PBS were injected using a U-100 insulin syringe (BD Micro-Fine) in C57Bl/6J mice. Some 1 × 106 cells OSCC13-LUC (luciferase expression vector expressing cells) were similarly grafted. Tumors were visible 2 weeks upon engraftment and mice were sacrificed for analysis at 3 weeks (bioluminescence experiment), 2 and 4 weeks (NIR) or 4 weeks (IR). For irradiation analysis, a 2 Gy unique dose of photons was delivered two weeks after tumor cell engraftment (3 × 106 cells). Mice were sacrificed two weeks later and tissue was analyzed by staining. For bioluminescence detection, a RediJect D-Luciferin Ultra Bioluminescent Substrate (Perkinelmer 770505) solution at 30 mg/ml was injected intraperitoneally 7 min before imaging. Images were acquired for 5 min using a live imager (NightOwl, Berthold). All mice were housed and handled according to the guidelines of INSERM and the ethical committee of Alsace, France (CREMEAS) (Directive 2010/63/EU, 01386.02, C-67-482-033 on the protection of animals used for scientific purposes).
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4

Bioluminescence Imaging of Rats under Isoflurane

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Bioluminescence imaging of rats was performed under 3% inhaled isoflurane anesthesia. Bioluminescence imaging used the IVIS Lumina II (PerkinElmer) (L‐ECT group, n=6; ECT group, n=6) for imaging. Ten minutes before the measurement of luminescence, 150 mg/kg body weight of Rediject D‐luciferin Ultra Bioluminescent Substrate (PerkinElmer) was injected intraperitoneally. We used an integration time of 1 minute, where images were acquired every 2 minutes until observing a steady decline in signal intensity. All images were analyzed with Living Image Software (PerkinElmer).
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5

In Vivo Bioluminescent Imaging of Mice

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Prior to imaging, the mice were i.p. injected with 100 μL of 30 mg/ml RediJect D-Luciferin Ultra bioluminescent substrate (PerkinElmer). Mice were then anesthetized with isoflurane and imaged within 20 minutes for bioluminescence using the in vivo imager. All the bioluminescence data, except for those shown in Supplementary Figures 7d and e, were collected using the IVIS Spectrum In Vivo Imaging System (PerkinElmer), and bioluminescent photon outputs were quantified using the Living Image Software (PerkinElmer). The bioluminescence data shown in Supplementary Figures 7d and e were collected using the Lago X optical imaging system (Spectral Instruments Imaging) and analyzed using Aura software (Spectral Instruments Imaging).
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6

Establishing mir-155 Lymphoma Xenografts

Check if the same lab product or an alternative is used in the 5 most similar protocols
To establish mir-155LSLtTA lymphoma subcutaneous flank tumors, first enlarged spleens were extracted from 2–3 month-old mir-155LSLtTA mice with obvious lymphadenopathy (which generally correlated with incidence of splenomegaly). Using a 100 μm pore size cell strainer technique, spleen tissue was dispersed into a single cell suspension in 5% FBS in PBS on ice. Red blood cells were lysed using ammonium chloride lysis buffer (Stem Cell Technologies), and 5 × 106 cells were subcutaneously injected into nude mice. Tumors were generally palpable within 10 days; tumor volume was calculated as (Length x Width2)/2.
For bioluminescent xenograft tumors, KB cells were stably transfected with firefly luciferase and clonally selected via hygromycin B selection; 5 × 106 cells were subcutaneously injected into nude mice to establish tumors. RediJect D-Luciferin Ultra Bioluminescent Substrate (PerkinElmer) was administered via the manufacturer’s protocol for intravital monitoring of tumor bioluminescence using IVIS Spectrum (Caliper). It was pre-established that for all flank tumor studies, animals were excluded if their tumors had not reached a volume of 50–100 cm3 by the time of treatment. Animals were randomized into experimental arms by minimizing the differences in mean tumor size and standard deviation.
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