BLI was used to follow in vivo growth of Fluc- or Rluc-engineered implanted tumor cells over time using a PerkinElmer IVIS Lumina system. For Fluc imaging, mice were imaged 7 min after intraperitoneal injection of d -luciferin (#122799, PerkinElmer). For Rluc imaging, mice were imaged 1 min after intravenous injection of coelenterazine h (#760506, PerkinElmer).
Coelenterazine h
Manufactured by PerkinElmer
Coelenterazine h is a bioluminescent substrate used in various laboratory applications. It is a core component in the detection and visualization of biological processes involving luciferase enzymes. The compound produces a luminescent signal when oxidized, enabling real-time monitoring and quantification of cellular activities and interactions.
Automatically generated - may contain errors
Lab products found in correlation
D luciferin, by PerkinElmer (2 mentions)
Ivis lumina system, by PerkinElmer (1 mentions)
Ivis lumina 3, by PerkinElmer (1 mentions)
Centro lb 960, by Berthold Technologies (1 mentions)
Dual luciferase reporter assay kit, by Promega (1 mentions)
Protein assay, by Bio-Rad (1 mentions)
Caspase glo 3 7, by Promega (1 mentions)
Glomax, by Promega (1 mentions)
Celltiter glo, by Promega (1 mentions)
Ufc901024, by Merck Group (1 mentions)
Living image 2, by PerkinElmer (1 mentions)
D luciferin, by Promega (1 mentions)
4 protocols using coelenterazine h
1
In vivo Bioluminescence Imaging
2
In vivo and in vitro Renilla Luciferase Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In vivo, Renilla luciferase activity was measured in the TA, quadriceps and gastrocnemius muscles of p16-3MR mice. Anaesthetized mice were injected intramuscularly with coelenterazine H (PerkinElmer, 760506) and luciferase activity was immediately measured using the IVIS Lumina III (PerkinElmer) system. In vitro, Renilla luciferase activity was measured from the cryopreserved diaphragm and TA muscles using the Dual-Luciferase Reporter Assay Kit (Promega, E1910). The signal was measured using the luminometer Centro LB 960 (Berthold Technologies) and values were normalized to the total protein extracted measured using Bradford method (Protein Assay, Bio-Rad, 500–0006), and the damaged area was measured after haematoxylin and eosin (H&E) staining.
3
Evaluation of Anti-EGFR Protein Therapies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumor cells were plated in 96-well plates and treated with different doses of anti-EGFR VHH (EV), DR ligand (DRL), EVDRL, or anti-EGFR scFv-TRAIL (ESDRL) and control media for 24, 48, and 72 hours. To obtain conditioned media containing these proteins, lentiviral plasmid vectors coding for EV, DRL, EVDRL, and ESDRL were transfected into 293T cells. Medium was changed the next day, collected 40 hours after transfection, concentrated using centrifugal filter (#UFC901024, MilliporeSigma), and stored at −80°C until future use. Their concentrations were quantified by enzyme-linked immunosorbent assay (ELISA). Control media were made from GFP control–transduced cells transduced in parallel with EV, DRL, EVDRL, and ESDRL. Cell viability was measured using an adenosine triphosphate–dependent luminescent reagent (CellTiter-Glo, #G755A, Promega; Glomax, Promega) according to the manufacturer’s instructions for non-Fluc–expressing cells or with d -luciferin (#122799, PerkinElmer) and coelenterazine h (#760506, PerkinElmer) for Fluc- and Rluc-expressing cells, respectively. Caspase-3/7 activity was determined using a DEVD-aminoluciferin assay (Caspase-Glo 3/7, #G8091, Promega) according to the manufacturer’s instructions. All experiments were performed in triplicates.
4
Bioluminescent Imaging of Stem Cells and Tumors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were anesthetized with isoflurane (2% induction, 1.5% maintenance) and placed in the IVIS 100 optical imaging system (Perkin Elmer), maintaining the body temperature at 37°C. Subsequently, D-luciferin (126 mg/kg, Promega) or coelenterazine-h (15 μg/mouse, Perkin Elmer) were intravenously injected to assess stem cell or hU87 tumor cell viability, respectively. Subsequently, bioluminescent images were acquired. Data were analyzed for maximum intensity of the photon flux by the living image 2.50.1 software (Perkin Elmer).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!