Babco, Signet, Sternberger Monoclonals, Senetek, Covance

other

Helix NP™ NIR

425301

1 mL

265 USD

425301

Helix NP™ NIR

1 mL

265 USD

NONAB_CHMPRB

Helix NP™ NIR is a cell-impermeant nucleic acid probe suitable for use as a viability dye in flow cytometry. It can also be used for viability in microscopy on live cells or as a nuclear counterstain on fixed and permeabilized cells and tissues. It is a far-red emitting dye with an excitation/emission max of 640 nm/660 nm that can be detected in the Alexa Fluor® 647 or APC channel.Protocol for flow cytometric viability staining using Helix NP™ NIR:;Isolate cells following protocol of choice.;Optional For multicolor flow cytometry experiments, surface-stain cells as recommended. Helix NP™ NIR should be added as the last step prior to sample acquisition.Note: The use of Helix NP for viability staining is incompatible with cell fixation and permeabilization protocols. If cells are to be fixed and permeabilized, use a fixable viability dye, such as a Zombie™ Fixable Viability kit.;Dilute Helix NP NIR to required concentration. We have observed good flow cytometric viability staining in a final concentration of 5 - 50 nM Helix NP™ NIR, but recommend titrating the reagent to determine the optimal concentration for cells of interest.For example, to stain cells in a final concentration of 50 nM Helix NP NIR, prepare a 1:2000 dilution of the 1.0 mM stock in Cell Staining Buffer (or equivalent). Then, add 50 µL of diluted reagent to 450 µL of cell suspension.;Do not wash cells after adding Helix NP™ NIR. Samples are ready for acquisition.;Analyze cells on a cytometer equipped with a 633 red laser.;Protocol for nuclear counterstaining fixed and permeabilized cell specimens using Helix NP™ NIR:;Fix cultured cells with 1% - 4% Paraformaldehyde (PFA) for 10 minutes at room temperature.;Wash the cells two times with 1X PBS.;Permeabilize the cells with 0.5% Triton X-100 for 10 minutes at room temperature.;Wash the cells two times with 1X PBS.;Block cells with 5% fetal bovine serum for 30 minutes at room temperature.;Prepare the working solution.We recommend titrating the reagent to determine optimal concentration for cells of interest. We have observed good results in the 0.5 - 5 µM range for IHC-F and IF/ICC;Stain the cells with diluted solution for 20 minutes at 4°C or room temperature in dark.Protect from light prior to imaging.;Wash the cells twice with 1X PBS.;Incubate cells with 1X PBS for 10 minutes in 4°C in dark.;Mount the slides with a media and image the slides.;