Click-iT™ EDU Zellproliferationskit für Bildgebung, Alexa Fluor™ 647 Farbstoff

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.

The problem is likely not the Alexa Fluor 594 azide. Since there are no alkynes endogenous to mouse tissue, there is nothing for the dye-azide to bind to. Since the background doesn't overlap with nuclei (DAPI signal), this isn't an issue of unintended EdU labeling. This red is autofluorescence from red blood cells; they autofluoresce in the red and don't have nuclei. This can be confirmed by checking a completely unlabeled tissue section (no dye present at all) to see if they are still present and by examining the cells at high magnification and looking for corpuscular shape.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Yes. You can purchase Alexa Fluor 647 Azide, Triethylammonium Salt (Cat. No. A10277). Unfortunately, the actual concentration of the azide dyes used in the Click-iT assay kits is proprietary. However, we recommend making up a 1-10 mM azide stock solution in high-quality anhydrous DMSO and using it at approximately 1-5 µM final concentration in the click reaction. You will need to work out a dye concentration empirically. Too high of a dye concentration can result in high non-specific background or dye aggregates, whereas too low of a concentration may not give a strong enough signal.

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

We do not recommend using phalloidin conjugates for staining actin in combination with traditional Click-iT or Click-iT Plus reactions since phalloidin is extremely sensitive to the presence of copper.

For staining actin in combination with traditional Click-iT or Click-iT Plus reactions, we recommend using anti-α-actin antibodies for staining actin in the cytoskeleton. You can find a list of our actin antibodies here.

Another option would be to use the Click-iT Plus Alexa Fluor Picolyl Azide Toolkit (Cat. Nos. C10641, C10642, C10643). These Click-iT Plus toolkits provide Copper and Copper protectant separately which makes it easier to titrate the copper concentration to obtain optimal labeling with minimal copper-mediated damage. You may need to optimize the click reaction with the lowest possible concentration of copper and then perform the phalloidin staining.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Yes, but the standalone products are not shipped at the same amount as provided in the Click-iT EdU kits; the amount of dye-azide provided in the Click-iT kits is proprietary information. See these catalog numbers for the stand alone products:
- Cat. No. A10266: Alexa Fluor 488 azide
- Cat. No. A10270: Alexa Fluor 594 azide
- Cat. No. A10277: Alexa Fluor 647 azide

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.