What color is Alexa Fluor 647?
far-red
Invitrogen Alexa Fluor 647 dye is a bright, far-red–fluorescent dye with excitation ideally suited for the 594 nm or 633 nm laser lines.
What laser is PE on?
PE is also excitable by the 561 nm laser and so PE based fluorophores appear on both lists. However, as PE is maximally excited at 546 nm, where possible it is advisable to use the 561 nm laser to get the maximal signal and to reduce spillover and therefore compensation.
Can you use PerCP Cy5 5 with APC?
For example, PE-Cy™5 is very bright with regard to stain index (Table 1), but it has considerable spillover (ie, adds background) into the APC detector. While these two fluorochromes can be used together, the resolution sensitivity in APC will be reduced compared to, for example, a combination of PerCP-Cy5. 5 and APC.
Can I use APC and APC Cy7 together?
In my experience, it is not advisable to use APC/Cy7 and PE/Cy7 together due to heavy cross-beam contamination as their emission max are exactly same. However, they can be used together if you are using machines like the Cytek Aurora.
Is 6-FAM the same as FAM?
Commercially available FAM is a mixture of two isomers, 5-FAM and 6-FAM, and the correct name is 5(6)-carboxyfluorescein. The dyes are membrane-impermeant and can be loaded into cells by microinjection or scrape loading.
Can PerCP and APC be used together?
While these two fluorochromes can be used together, the resolution sensitivity in APC will be reduced compared to, for example, a combination of PerCP-Cy5. 5 and APC.
Is Cy5 a protein?
The Cy5-labeled protein has excitation and emission wavelengths at 650 nm and 670 nm respectively, and can be directly used for downstream applications including ELISA, western blot, Immunohistochemistry, Immunofluorescence, and FACS, etc.
Can you reuse secondary antibody?
Finally, very few researchers will reuse secondary antibodies, as you generally burn through fewer of them per equivalent volume. Secondary antibodies are also cheaper and used at far more dilute ranges. The decision to reuse antibodies overall is largely project dependent.
What is the difference between Alexa Fluor 647 and Cy3?
As N-hydroxysuccinimidyl ester dyes, the Alexa Fluor 555 dye was similar to the Cy3 dye, and the Alexa Fluor 647 dye was similar to the Cy5 dye with respect to absorption maxima, emission maxima, Stokes shifts, and extinction coefficients.
What is the maleimide derivative of Alexa Fluor 647?
The maleimide derivative of Alexa Fluor® 647 is the most popular tool for conjugating the dye to a thiol group on a protein, oligonucleotide thiophosphate, or low molecular weight ligand. The resulting Alexa Fluor® 647 conjugates exhibit brighter fluorescence and greater photostability than the conjugates of other spectrally similar fluorophores.
What is Alexa Fluor 647 used for?
Alexa Fluor® 647 is a bright, far-red fluorescent dye with excitation ideally suited for the 594 nm or 633 nm laser lines. Used for stable signal generation in imaging and flow cytometry, Alexa Fluor® 647 dye is water soluble and pH-insensitive from pH 4 to pH 10.
Can I replace conventional fluorophores with Alexa Fluor dyes?
Conventional fluorophores and their conjugates can often be replaced with spectrally similar Alexa Fluor dyes and conjugates without affecting optical filter choices or other instrumentation considerations ( Table 2 ).