5 replies to this topic

[soymilk14]

i will do immunofluorescence, i want to ask if i can use dsred staining for one protein and  alexa fluor 488 in another protein. Which fluorophore is the best to use together with dsred? thanks

[laurequillo]

Well, actually DsRed (if I am not mistaken) is a fluorescent protein itself, not a dye, so yeah sure, you can transfect your protein fused to DsRed and then use an antibody against other protein and use the alexa 488 as secondary (it is green). So you will have one protein in red, and the other one in green.

[soymilk14]

thank you. i still have additional question...

if i have a proteinfused with gfp,cfp and dsred and i want to check the proliferation using brdu-fitc, will it be possible?

[laurequillo]

soymilk14, on Jul 7 2010, 06:38 PM, said:

thank you. i still have additional question...

if i have a proteinfused with gfp,cfp and dsred and i want to check the proliferation using brdu-fitc, will it be possible?

Actually i am not sure... The gfp you can use, but the dsred I guess use the same channel that you use for the fitc

[Rsm]

FITC is in the same channel as GFP, not dsRed. So you can't use BrdU-FITC and GFP together.

[laurequillo]

Minna, on Jul 14 2010, 10:36 AM, said:

FITC is in the same channel as GFP, not dsRed. So you can't use BrdU-FITC and GFP together.

that is right, sorry, I wrote it in the other way around!

Edited by laurequillo, 14 July 2010 - 12:51 AM.