Akalumine hcl

Manufactured by Fujifilm

Sourced in Japan

AkaLumine-HCl is a chemical compound used in various laboratory applications. It serves as a fluorescent labeling agent, enabling the detection and visualization of biomolecules and other materials. The core function of AkaLumine-HCl is to provide a reliable and sensitive fluorescent signal for analytical and research purposes.

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Lab products found in correlation

Ivis imager, by PerkinElmer (1 mentions) Black 96 well plate, by PerkinElmer (1 mentions) Fluorofurimazine ffz, by Promega (1 mentions) Victor nivo multi plate reader, by PerkinElmer (1 mentions) Nano glo luciferase assay system, by Promega (1 mentions) Clariostar plate reader, by BMG Labtech (1 mentions)

4 protocols using akalumine hcl

Bioluminescence Imaging of SARS-CoV-2 Infection

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Luminescence was detected using AkaLumine-HCl (FUJIFILM Wako Chemicals), and luciferase activity in vitro was measured by the 10-s integral analysis using an AB-2270 Luminescencer Octa (Atto) according to the manufacturer’s protocol. VeroE6/TMPRSS2 cells were seeded in a black 96-well plate (PerkinElmer) at a density of 15,000 cells per well. Serially diluted B.1.1, B.1.1-NanoLuc, or B.1.1-Akaluc were used to infect the cells. At 32 hpi, cells were washed once with PBS and imaged using an IVIS Imager (PerkinElmer) at 5 min after addition of substrate: fluorofurimazine (FFz) (3 g/mL: Promega) for NanoLuc and AkaLumine-HCl (4.3 g/mL) for Akaluc. The following conditions were used for image acquisition: open emission filter, exposure time = 3 s, binning = medium: 8, field of view = 13.2 × 13.2 cm and f/stop = 1.
For in vivo experiments, infected hamsters or mice were injected with FFz (440 nmol/g) or AkaLumine-HCl (75 nmol/g) intraperitoneally according to the indicated protocol from the previous studies¹¹ (link)^,¹² (link) and the luminescence images were monitored by IVIS at 1, 2, and 3 dpi. The following conditions were used for image acquisition: open emission filter, exposure time = 60 s, binning = medium: 8, field of view = 13.2 × 13.2 cm and f/stop = 1. All images were analyzed using Living Image Ver4.7.3 (PerkinElmer).

Tamura T., Ito H., Torii S., Wang L., Suzuki R., Tsujino S., Kamiyama A., Oda Y., Tsuda M., Morioka Y., Suzuki S., Shirakawa K., Sato K., Yoshimatsu K., Matsuura Y., Iwano S., Tanaka S, & Fukuhara T. (2024). Akaluc bioluminescence offers superior sensitivity to track in vivo dynamics of SARS-CoV-2 infection. iScience, 27(5), 109647.

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Bioluminescent Reporter Assay Optimization

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Cells and cellular debris were removed from the culture medium by centrifugation at 2 000 × g at 4 °C for 10 min, and 50 μL of the supernatant was transferred to white-walled 96-well plates. Nano-Glo substrate (50 μL; FRZ) diluted at 1:50 (v/v) in the buffer provided with the Nano-Glo Luciferase Assay System (Promega) was added to the plates and the luciferase intensity was measured immediately using a VICTOR Nivo Multiplate Reader (PerkinElmer). For spectral scanning, a CLARIOstar plate reader (BMG Labtech, Aylesbury, UK) was used. To assess DTZ, another substrate of Antares2, 50 μL of DTZ (HY-111382; MedChemExpress, Monmouth Junction, NJ, USA) at the indicated concentrations was added to 50 μL of supernatant. To assess Akaluc, Akalumine-HCl (150 μM; FUJIFILM Wako chemicals, Osaka, Japan) was added to 50 μL of supernatant.

Hikita T., Miyata M., Watanabe R, & Oneyama C. (2020). In vivo imaging of long-term accumulation of cancer-derived exosomes using a BRET-based reporter. Scientific Reports, 10, 16616.

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Circadian Bioluminescence Monitoring in Reporter Cells

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The bioluminescence rhythm of spot cultures of reporter cells in white 96-well plates was monitored as described previously [25 (link)] except for the experiment in Fig 8B where we used spot cultures in black 24-well plates and a bioluminescence monitoring apparatus CL24-LIC (Churitsu Electric Corp., Nagoya, Japan). The circadian parameters (period, phase, and amplitude) were calculated by the cosinor method of the Rhythm Analyzing Program [54 (link)]. The amplitude was estimated from the overall average best-fitting cosinor curves between the second and the fifth day of bioluminescence monitoring. To monitor the clock protein-luciferase fusion reporters (Figs 2D, 6 and 7 and S3 Fig), we used AkaLumine-HCl (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) [55 (link)] as the substrate because the bioluminescence rhythm was more stable against environmental disturbances (e.g., light on/off, drying of agar, vibration of culture plates).

Matsuo T., Iida T., Ohmura A., Gururaj M., Kato D., Mutoh R., Ihara K, & Ishiura M. (2020). The role of ROC75 as a daytime component of the circadian oscillator in Chlamydomonas reinhardtii. PLoS Genetics, 16(6), e1008814.

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Bioluminescent Murine Model for Cancer

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All studies involving animals were performed in accordance with the institutional guidelines for the use of laboratory animals and approved by the Review Board for Animal Experiments of the University of Tokyo (approval ID PA18‐42). NOD.Cg‐Prkdc^scid Il2rg^tm1Sug Tg (SRa‐IL3, CSF2)/Jic (NOG IL‐3/GM‐Tg) mice expressing human IL‐3 and GM‐CSF were purchased from the Central Institute for Experimental Animals (Kawasaki Japan).¹⁸ MDS‐L/Akaluc cells (1 × 10⁷ cells) were inoculated into female NOG IL‐3/GM‐TG mice irradiated at a dose of 1.8 Gy. Three weeks after tumor inoculation, mice were randomly divided into three groups (6 mice per group) and treated with the indicated compounds. Just before the imaging analysis, 100 µL of 5 mM AkaLumine‐HCl (Wako) was injected intraperitoneally into mice, and mice under isoflurane anesthesia were imaged within 5–10 minutes of the injection. The following conditions were used for image acquisition: open for total bioluminescence, exposure time = 60 seconds, binning = 4‐8, field of view = 25 × 25 cm, and f/stop = 1. In vivo photon counting was conducted with an IVIS system using Living Image 2.5 software (Xenogen). Mice were monitored until they became moribund, at which time they were killed.

Zhong C., Kayamori K., Koide S., Shinoda D., Oshima M., Nakajima‐Takagi Y., Nagai Y., Mimura N., Sakaida E., Yamazaki S., Iwano S., Miyawaki A., Ito R., Tohyama K., Yamaguchi K., Furukawa Y., Lennox W., Sheedy J., Weetall M, & Iwama A. (2020). Efficacy of the novel tubulin polymerization inhibitor PTC‐028 for myelodysplastic syndrome. Cancer Science, 111(12), 4336-4347.

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