Click-iT™ Plus OPP Alexa Fluor™ 647 Protein Synthesis Assay Kit

Q: Can the Click-iT Plus OPP Alexa Fluor 647 Protein Synthesis Assay Kit (Cat. No. C10458) be used for detection in flow cytometry?

The Click-iT OPP Assay Kits (Cat no. C10456, C10457 and C10458) can be used with any application that detects whole cells; therefore, the assay can be adapted for flow cytometry. You can follow the imaging protocol for labelling in the user guide. Briefly, you would need to spin the cells down (<2,000 rpm) to wash instead of covering the cells with the Click-iT Reaction Rinse Buffer as mentioned in step 5.6 on page 7 of the user guide. The picolyl azide dye concentration used in the click reaction has been optimized for imaging, which is less sensitive than flow. We recommend reducing this volume from 1/5th up to 1/10th the protocol volume for flow detection and making up the difference with 1X Reaction Buffer while keeping the volumes of the click reagents the same as in the protocol. Find additional tips, troubleshooting help, and resources within our Labeling Chemistry Support Center .

Click-iT™ Plus OPP Alexa Fluor™ 647 Protein Synthesis Assay Kit - FAQs

2 product FAQs found

I will be performing a cell proliferation assay using Click-iT EdU kit. At what point can I stop overnight, or do I have to perform all the steps continuously?

One may store the sample after fixation overnight in PBS at 4^oC. For longer storage (<1 week) , store in buffer with 1-2% formaldehyde or in formalin to limit microbial growth. If you use sodium azide as a microbial inhibitor, it must be completely removed prior to the Click-iT reaction.

Find additional tips, troubleshooting help, and resources within our Cell Viability, Proliferation, Cryopreservation, and Apoptosis Support Center.

Can the Click-iT Plus OPP Alexa Fluor 647 Protein Synthesis Assay Kit (Cat. No. C10458) be used for detection in flow cytometry?

The Click-iT OPP Assay Kits (Cat no. C10456, C10457 and C10458) can be used with any application that detects whole cells; therefore, the assay can be adapted for flow cytometry. You can follow the imaging protocol for labelling in the user guide. Briefly, you would need to spin the cells down (<2,000 rpm) to wash instead of covering the cells with the Click-iT Reaction Rinse Buffer as mentioned in step 5.6 on page 7 of the user guide. The picolyl azide dye concentration used in the click reaction has been optimized for imaging, which is less sensitive than flow. We recommend reducing this volume from 1/5th up to 1/10th the protocol volume for flow detection and making up the difference with 1X Reaction Buffer while keeping the volumes of the click reagents the same as in the protocol.

Find additional tips, troubleshooting help, and resources within our Labeling Chemistry Support Center.