Babco, Signet, Sternberger Monoclonals, Senetek, Covance

other

Helix NP™ Green

425303

250 μL

211 USD

425303

Helix NP™ Green

250 μL

211 USD

NONAB_CHMPRB

Helix NP™ Green 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 green-emitting dye with an excitation/emission max of 495 nm/519 nm that can be detected in the Alexa Fluor® 488 or FITC channel.Protocol for flow cytometric viability staining using Helix NP™ Green:;Isolate cells following protocol of choice.;Optional For multicolor flow cytometry experiments, surface-stain cells as recommended. Helix NP Green 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 Green to required concentration. We have observed good flow cytometric viability staining in a final concentration of 1 - 10 nM, but recommend titrating the reagent to determine optimal concentration for cells of interest.For example, to stain cells in a final concentration of 10 nM Helix NP Green, prepare a 1:5000 dilution of the 5.0 mM stock in Cell Staining buffer (or equivalent). Then, add 5 µL of diluted reagent to 500 µL of cell suspension.;Do not wash the cells after adding Helix NP™ Green. Samples are ready for acquisition.;Analyze cells on a cytometer equipped with a 488 nm blue laser.;Protocol for nuclear counterstaining fixed and permeabilized cell /tissue specimens using Helix NP™ Green:;Fix specimens with 1%-4% Paraformaldehyde (PFA) for 10 minutes at room temperature.1% for cultured cells;4% for frozen tissue;Wash the cells/tissue two times with 1X PBS.;Permeabilize the cells/tissue with 0.5% Triton X-100 for 10 minutes at room temperature.;Wash the cells/tissue two times with 1X PBS.;Block cells with 5% fetal bovine serum for 30 minutes at room temperature.;Complete any additional blocking steps, antibody staining, and washes before adding the Helix NP™ Green counterstain.;Prepare the working solution.It is best to try a range of dye concentrations to determine the optimal concentration for cells/tissues of interest to achieve the best image. We have obtained good results using a range of 0.5 - 10 µM for IHC-F and IF/ICC).;Stain the cells/tissues with the working solution for 20 minutes at 4°C or room temperature in the dark.Protect from light prior to imaging.;No wash step is needed after staining.;Mount the slides with an antifade medium.;Image slides with a filter ideal for ex/em max of 495 nm/519 nm (Alexa Fluor® 488 or FITC channel.;