<br><font size=2 face="sans-serif">Hello Manuelle</font>
<br>
<br><font size=2 face="sans-serif">may I ask you to check your secondary antibody? How does the control look like without applying the primary ? </font>
<br><font size=2 face="sans-serif">We use both for CD4 and for CD8a on mouse frozens antibodies( host rat) from Pharmigen. As secondary I use a rabbit anti-rat biotinylated one, <i>mouse adsorbed</i> from Vector.</font>
<br><font size=2 face="sans-serif">For the blocking of endogenous peroxidase don't use methanol (!) , it weakens the signal in most cases. For the unspecific binding block we use 10% normal rabbit serum (-> secondary host species).</font>
<br><font size=2 face="sans-serif">I hope this helps.</font>
<br><font size=2 face="sans-serif">Good luck,</font>
<br>
<br><font size=2 face="sans-serif">Antje Marcantonio<br>
Novartis Pharma AG<br>
BU Transplantation Research<br>
Basel, Switzerland<br>
Tel. +41 61 324 6730<br>
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