[Histonet] immunofluorescence staining in frozen tissue

[Kelly D Mcqueeney]

I used Ki67 antibody (ab833 at 1:50) from Abcam or H-300 antibosy at 
1:200 from Santa Cruz. I fix the tissue in ice-cold acetone for 5 
minutes. I found it was very important to permeabilize the tissue with 
TBS + 0.025% TritonX-100 (follow Abcam protocol) for 5 minutes before 
blocking. If I deleted this step, the nuclear staining was very weak for 
either antibody..

Best of luck,
Kelly

Kelly Yang wrote:

>Dear histonetters,
>
> 
>
>We are working on immunofluorescence staining with two markers, ki67 and DNMT1, in human bladder tissue. I had tried ki67 from Dako and DNMT1 from Imgenex, but none of them give us the good result. 
>
> 
>
>Does anyone have a protocol for staining frozen tissue with Ki67 or DNMT1? Also which antibody are you recommending? 
>
>Thank you for your help in advance. 
>
> 
>
>
>
>
>Kelly Yang
>Graduate student 
>Department of Epidemiology
>School of Public Heath
>University of California, Los Angeles
>310-409-9179
>ext. 57795
>yangyc <@t> ucla.edu
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