[Histonet] CD168 (2D6) antibody

[Haviland,Joie]

Hello...
I am attempting to use this antibody, CD168 (2D6), also known as Rhamm or Hmmr.

I tried using 10mM citrate buffer but that did not work at all. I did not see any signal. I also have tried using the 1mMEDTA antigen retrieval solution but the control tissue, normal human testis, was no longer there after AR is done. I have tried other control tissues still no results. It was suggested to me to use Excel slides which I did and the tissue stayed but was very degraded after AR.

 I am using a 1:50 and 1:100 dilution of this antibody but I see a bit of cytoplasm lighting up at 1:50. The data sheet says that it should show on the cell membrane.   But the signal only showed up after I left the antibody overnight at 4 degrees.  I have also tried the antibody at room temp for 1 hour to no avail.  I am using DAKO products, protien block, Mouse Polymer and DAB+ which have worked in other IHC's recently done.

I have tried two different lots of this antibody with no success.  Any suggestions?

Thanks
Joie
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Joie Haviland, HT (ASCP)
Senior Research Assistant,
Dr. McDonnell's Lab Y5.6057
Department of Hematopathology
MD Anderson Cancer Research Center
713-792-0705