[Histonet] 33D1 DC marker

[koellingr <@t> comcast.net]

Sonya,
What were the frozen sections of tissue-wise?
33D1 is a fairly recent and I've used it to stain in the IHC configuration you tried but remember that that target is not at all well characterized.  It certainly doesn't appear to be a structural or housekeeping protein expressed in abundance.
1) Look at flow diagrams of something like spleen stained 33D1 and that 1 log shift in "dendritic cells?" is a really minute portion of the population and flow people are looking at 5-10K gated events (cells).  It is possible that depending on what tissue you section and how big it is, there just may be very few 33D1+ cells around to see.
2) Again, the jury is out on what 33D1 even is.  Certainly it can be upregulated or downregulated (see articles on GM-CSF stimulation or IL-4 induction of down-regulation available in literature).
3) How were your coverslip DC's isolated?  Certainly they cannot, after multiple sorts or purifications,  be very similar to in-vivo DC's.
4) to be sure to get enough to see, I'd do in-vivo stimulation of the mouse.  There is literature on how to do that.
5) there are other companies that sell 33D1, if you are convinced BD (that I think is great) 33D1 is not working.

Ray
unemployed and at home so no current lab affiliation to list

-------------- Original message -------------- 
From: "Martin S." <sonya.martin <@t> soton.ac.uk> 

> Hi All, 
> 
> Has anyone used 33D1 (mouse DC marker) antibody from BD Pharmingen on 
> frozen sections? I've tried it with the secondary they recommend (mouse 
> anti-rat IgG 2b) but I dont see anything. 
> I tried it on isolated DC's growing on coverslips and got a very faint 
> staining. 
> 
> Any comments? 
> 
> Sonya 
> 
> 
> 
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