FAQ  •  Register  •  Login

CCR7 erased

Forum rules
It is fine to promote your company's reagents. Just make sure they are relevant to CyTOF, and do so in moderation and style :-)
<<

mayanlevy

Participant

Posts: 9

Joined: Thu May 11, 2017 10:26 am

Post Thu Oct 04, 2018 5:06 pm

CCR7 erased

Hello,

We seem to have a problem with the CCR7-159Tb antibody from Fluidigm (Clone G043H7) - although the antibody should be enough for 50 tests, 1ul is not enough.
Our protocol is as follows: Barcoding, EC staining, fix, Methanol perforation, IC staining, fix, Ir staining.
All the other antibodies work well, and only the CCR7 is not. We have done a calibration experiment where we found only using 5ul of the antibody worked. Using different vendors with 5ul volume of the antibody did not yield a good result...
Fluidigm claims the problem is with the barcoding fix.
We need to have both the barcoding and methanol perm for pFlow, and at the same time CCR7 is imporatant to our panel.
Did anyone run into a similar problem? If so, what was your solution?
Thanks.
<<

nlazarus

Contributor

Posts: 20

Joined: Wed Nov 13, 2013 7:28 pm

Post Thu Oct 04, 2018 7:10 pm

Re: CCR7 erased

You could pre-stain with CCR7 and any other sensitive markers prior to the BC fixation. After staining and washing, proceed as you normally would. I have done that with success.

Hope that helps.

Nicole

Return to Reagents for CyTOF

Who is online

Users browsing this forum: No registered users and 8 guests