Biolegend, 425303, Helix NP™ Green
250microl
Catalog Number: 425303
Send an RFQ.
Category
Loading…
| Preparation | Helix NP™ Green is supplied at 250 µl per vial. |
|---|---|
| Concentration | 5.0 mM |
| Storage & Handling | Upon receipt, store at -20°C. |
| Application | FC - Quality tested IF, IHC-F - Verified |
| Recommended Usage | For use in viability assays, the optimal concentration can range from 1 - 10 nM. For immunofluorescence microscopy and immunohistochemical staining on frozen tissue sections, the suggested use of this reagent is 0.5 - 10 µM. It is recommended that the reagent be titrated for optimal performance for each application. |
Unless otherwise stated, the minimum order amount is USD 1,000.
For orders below USD 1,000, an additional handling fee may apply, or we may arrange local delivery / local handover where available.
All quotations are subject to final confirmation, including product availability, lead time, batch information, shipping cost, and applicable export or handling requirements.
For quotation requests, please send an RFQ from website or contact:
Please send inquiries directly to us in the “To” field. Emails sent via BCC, mass mailing, or with unrelated recipients may be automatically filtered and will not be processed.
| Application Notes |
|---|
| 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. |
| Biology Area | Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Cycle/DNA Replication, Cell Proliferation and Viability |
|---|
| Gene ID | NA |
|---|
| Regulatory Status | RUO |
|---|
