Re: ISH of VEGFs and their receptors

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From:"Tony Henwood" <>, "Agripina Suarez" <>
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Dear  Agripina,
> Histonetters, help!!!
> I have been trying to do in situ hybridization of VEGFs  (A, B, C, D) 
> and their receptors (Flt-1, -4 and Flk-1) in formalin-fixed, paraffin-
> embedded normal human vascular tissues and transplant heart 
> vessels but still unsuccessful so far.

Hybridisation at room temp,
Remove formamide from the probe solution as well as the stringency 
After hybridisation, wash in 2xSCC for 5 min only (donot try high and 
low stringency washings).
Once you get some results, look at your technique and add the above 
steps only if your results are not specific.

Hope this helps,
Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital

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