CyTOForum

Hi Guys,

I have a new marker on my CyTOF panel that I believe is expressed "everywhere". It is cytoplasmic, secreted and nuclear.
I was wondering which protocol I should use for that.
My idea was to use the Maxpar Cytoplasmic/Secreted Antigen Staining with Fresh fix protocol from Fluidigm.
For that, I should use Maxpar Fix I Buffer (5x) cat#201065 and Maxpar Perm-S Buffer cat# 201066
I was wondering if I could use MAxpar solutions from Fluidigm Barcode for this.
In this case instead of Maxpar Perm-S Buffer I will be using Maxpar 10X Barcode Perm.
The Maxpar Fix I Buffer looks the same to me but they have different catalog numbers. The barcode kit FIX I solution is #S00115. Does anyone knows if they are the same?

Thanks very much!

All the Best!

Thais

Hi Thais,

I'm not sure I understand what you're asking.

If all that you're trying to do is detect the presence or absence of it, then surface staining is probably going to be the cleanest (assuming you have enough expression that you have a good signal intensity). In that case, you could stain fresh (live) cells before fix or perm.

Is there a *meaningful* difference between the localization of the protein within the cell? For example, there are some markers (particularly with transport or vacuoles) which *are* always present internally in the cell, but their presence or movement to the surface means something different than just whether it's present in the cell at all.


Mike

Hi Thais,
A bit delayed response here but I googled the same question and your post came up. I called Fluidigm and S00115 is the same as 201065. When the Maxpar Fix I Buffer (5X) comes in the 20-Plex Barcoding kit the bottle has "S00115" as catalog #. I called Fluidigm who confirmed.

What did you end up doing for your "everywhere" marker? Did the Cytoplasmic/Secreted Antigen Staining protocol work for the marker?

Emily