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Hypoxia Probe, LOX-1

Q.501 – What solvent should be used for the reagent?
DMSO is the recommended solvent.
Q.502 – Why the reagent sometimes doesn’t dissolve properly in old DMSO?
Old DMSO maybe contain moisture due to the absorbency in bad condition. We recommend using new one or the one stored in good condition, because it will be difficult for LOX-1 to dissolve into moisturized DMSO.
Q.503 – What is the storing condition or length of dissolved LOX-1 solution in DMSO?
LOX-1 solution is stable for couple of months under light protection in -20℃.
Q.504 – What is the recommended final concentration of LOX-1 for Hypoxia detecting assay?
We recommend adding LOX-1 into the culture medium at 2 µmol/L.
Q.505 – How can I detect/observe the signal/region of LOX-1?
One can detect the signal with microscope or cell imager that has fluorescence detector feature.
Q.506 – How long time is needed for hypoxia detection after LOX-1 treatment?
In monolayer culture, 2 hours is recommended. And in 3D culture, 24 hours is needed.
Q.507 – What type of filter is needed for detecting LOX-1 signal with fluorescent microscopy?
We have confirmed the adequateness of G-2A filter set (Nikon) and filter set 14 (Zeiss) for the observation. You can use the other filter set matching with the spectrum (Ex: 483, Em: 616) of LOX-1.
Q.508 – Can I measure the LOX-1 signal with microplate-readers
We don’t recommend that.
Q.509 – Is it possible to detect the LOX-1 signal with cell imagers?
Yes it is. We have confirmed that some cell imager equipment i.e. Celigo (Nexcelom), InCell series (GE Healthcare) and BioStation CT (Nikon) are capable to detect the signal.
Q.510 – Can I measure oxygen concentration accurately?
Measuring oxygen concentration can be possible by measuring phosphorescence lifespan. Because the phosphorescence lifespan is correlated with oxygen concentration.
Q.511 – Can I conduct time-lapse observation of the LOX-1 signal inside spheroid?
Yes, you can. But superoxide which is produced by excitation light maybe affect cytotoxicity to cells.
Q.512 – Can I use the reagent for cell culture under 2D condition?
Yes, you can.
Q.513 – Can LOX-1 visualize the hypoxic region of sliced spheroid or tissue?
No, it cannot. This is the reagent for only living cells.
Q.514 – Can I use the reagent for in vivo imaging?
Yes, you can. Please contact us for detailed information.
Q.515 – Are there any reference paper for detecting hypoxia of spheroids by LOX-1?
Yes, there are. Please see the paper list.