Aequorin photoprotein (untagged) is recombinantly produced in E. coli, purified via multi-step chromatography and appears as a yellow liquid.
Concentration
2 mg/ml in 1.2M (NH4)2SO4, 250 μl per tubeSequenceMTSKQYSVKLTSDFDNPRWIGRHKHMFNFLDVNHNGKISLDEMVYKASDIVINNLGATPEQAKRHKDAVEAFFGGAGMKYGVETDWPAYIEGWKKLATDELEKYAKNEPTLIRIWGDALFDIVDKDQNGAITLDEWKAYTKAAGIIQSSEDCEETFRVCDIDESGQLDVDEMTRQHLGFWYTMDPACEKLYGGAVP
Coelenterazine (native-CTZ) in sterile injection vials with a low retention volume. Each kit is supplied with two ‘Inject-A-Lume’ vials of 500 µg CTZ and 500 µl of sterile ‘Fuel-Inject’ diluent.
‘Fuel-Inject’ will improve the luciferase signal compared to conventional Ethanol-dissolved CTZ.
Recommended path of injection: intra-peritoneal cavity.
For more frequent injections we recommend our water-soluble Coelenterazine.
For animal use only.
Package Contents:500 ugs – 1 vial of 500 ugs lyophilized SOL-CTZ1 mgs – 2 vials of 500 ugs lyophilized SOL-CTZ10 mgs – 20 vials of 500 ugs lyophilized SOL-CTZ
Coelenterazine is often dissolved in Propyleneglycol (PG) in combination with other alcohols for in vivo imaging studies. Large intravenous doses of PG given over a short period of time can be toxic, including hyperosmolality, increased anion gap metabolic acidosis (due to lactic acidosis), acute kidney injury, and sepsis-like syndrome.
This can be avoided using water-soluble Coelenterazine. It is formulated to be isosmotic and easily secreted by the kidneys. Thus save for repeated i.v. injections. Water soluble Coelenterazine can be injected up to a 500 µg/100 µl concentration, achieving very high Coelenterazine levels within the body and leading to high Gaussia luciferase signals.
Application Example
Controls
We offer the solubilization vehicle without the Coelenterazine as injection control. Any side effects due to the injection process can be easily monitored, especially in sensitive applications like bioluminescent driven Optogenetics. One vial of the control (Cat. #3031-10+C) contains the same amount of solubilizer as one vial of Cat.#3031. The control will be dissolved in sterile water to create an iso-osmotic solution ready to inject.
To order control: please select ’10 mgs + 10 vials control’ under product options. This includes 20 vials of 500ug water-soluble CTZ and 10 vials of control.
Please inquire if larger quantities of Cat.#3031-10+C are needed.
Package Contents:500 ugs – 1 vial of 500 ugs lyophilized e-CTZ-SOL1 mgs – 2 vials of 500 ugs lyophilized e-CTZ-SOL10 mgs – 20 vials of 500 ugs lyophilized e-CTZ-SOL
e-Coelenterazine is often dissolved in Propyleneglycol (PG) in combination with other alcohols for in vivo imaging studies. Large intravenous doses of PG given over a short period of time can be toxic, including hyperosmolality, increased anion gap metabolic acidosis (due to lactic acidosis), acute kidney injury, and sepsis-like syndrome.
This can be avoided using water-soluble e-Coelenterazine. It is formulated to be isosmotic and easily secreted by the kidneys. Thus save for repeated i.v. injections. Water soluble e-Coelenterazine can be injected up to a 500 µg/100 µl concentration, achieving very high e-Coelenterazine levels within the body and leading to high Renilla luciferase signals.
Application Example
Controls
We offer the solubilization vehicle without the e-Coelenterazine as injection control. Any side effects due to the injection process can be easily monitored. One vial of the control (Cat. #3551-10+C) contains the same amount of solubilizer as one vial of Cat.#3551. The control will be dissolved in sterile water to create an iso-osmotic solution ready to inject.
To order control: please select ’10 mgs + 10 vials control’ under product options. This includes 20 vials of 500ug water-soluble CTZ and 10 vials of control.
Please inquire if larger quantities of Cat.#3551-10+C are needed.
50 ml NanoFuel® FLASH Reagent for Oplophorus Luciferase
1 ml 50X NLuc FLASH Substrate
Characteristics
one-step assay
bright luminescent signal for at least a few minutes
higher sensitivity than CTZ or Furimazine
low background, higher S/N ratio
enough reagent for 1000 assays
luminometer with injection port is recommended
although the substrate will diffuse into the cells this process can be accelerated by a lysis step, please select “add Lysis” under product options to add 50ml of5x Universal Lysisbuffer (CAT# 333) to your order
Graph below: HEK293 cells were transformed with NLuc and assayed 2 days later using a luminometer with injection port. Compared to other NLuc assay buffers, Cat. #324 increased the inital lightoutput with a reduced signal half-life. This data was kindly provided by the Neuroscience Department at Central Michigan University.
Aequorin photoprotein (untagged) is recombinantly produced in E. coli, purified via multi-step chromatography and appears as a white lyophilized powder.
Reconstitution
with Ca2+ free double distilled water
Sequence
MTSKQYSVKLTSDFDNPRWIGRHKHMFNFLDVNHNGKISLDEMVYKASDIVINNLGATPEQAKRHKDAVEAFFGGAGMKYGVETDWPAYIEGWKKLATDELEKYAKNEPTLIRIWGDALFDIVDKDQNGAITLDEWKAYTKAAGIIQSSEDCEETFRVCDIDESGQLDVDEMTRQHLGFWYTMDPACEKLYGGAVPApplicationpostitive control in Ca2+ assays
Coelenterazine (native-CTZ) in sterile injection vials with a low retention volume. Each kit is supplied with two ‘Inject-A-Lume’ vials of 500 µg CTZ and 500 µl of sterile ‘Fuel-Inject’ diluent.
‘Fuel-Inject’ will improve the luciferase signal compared to conventional Ethanol-dissolved CTZ.
Recommended path of injection: intra-peritoneal cavity.
For more frequent injections we recommend our water-soluble Coelenterazine.
For animal use only.
200 pUC18 Gluc sequence
