[Histonet] i need some technical help!

[Mr srinivas sv]

Hi,

Does anyone have a protocol or reference for cleaning up polyclonals
with acetone-extracted liver powder? 

I am trying to look at the expression of steroidogenic enzymes in the sheep ovary. My sections are cryosections. All my antibodies are polyclonal antibodies. My problem is that there is color development in both positive and negative controls. I am using normal rabbit sera (from SIGMA) for the negative control as my primary antibodies are raised in rabbit. I am using Normal goat sera and 1% BSA for blocking nonspecific antigens (Serum Blocking) as my secondary antibody is raised in Goat. I am also using 1% hydrogen peroxide for blocking endogenous peroxides. Fed up with this, i now want to try using acetone-extracted liver powder as an adsorbent. Can anyone explain me the use of acetone-extracted liver powder and protocol for using it for my experiment. 

I am trying this from past 6 months, without any positive result. So, anyone please troubleshoot my problem and suggest any alteration in my protocol. 

Please feel free to ask for any clarifications. 

Thanks... 

Regards, 

Srinivas




Srinivas Seekallu 
PhD student
Dept of Vet Biomed.Sci.
Western College of Veterinary Medicine,
University of Saskatchewan.
Saskatoon, Canada.

Ph: 306-477-0849 (Home)
    306-966-7373 (Lab)
    306-966-7382 (Office)
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