[Histonet] (no subject)

[shirin kouhpayeh]

 Dear histonetters
I'm working on a surface marker, uPAR, on lymph node tissue. For setting up the IHC-p procedure, I've chosen the ductal carcinoma sample as positive control. I use the citrate buffer ph=6 as retrieval solution and TBS ph=7.6 for wash steps. different Ag retrieval systems and different Ab ( santa cruz biotech. ) dilutions have been tested 
s 2*30 minutes of heating in water bath 70 degree of cellicius, 
5 minutes with 800 watt power in microwave oven following 15 minutes heating with power of 300
 6 minutes with max power ( 800 watt) following by 15 minutes on power 300 watt
10 minutes with max power following with 15 minutes on power 300 watt
20 min autoclave on 121 degree of cellicius 
But there is still no signal on ductal carcinoma sample. 
Is there anyone have a comment for me? 
I would be so appreciated .


Shirin kouhpayeh 
Ph.D student
Immunology dept. 
Isfahan university of medical science
Isfahan, Iran