Louise:
If that is the only thing you are dedicated to doing, you will have to make a literature search to find out how other people have handled this Ab: that would be one week, during which you can also locate archival tissues to test based in what has been done. If there is no published information this time will be reduced and added to the experimental phase.
Developing your "checker-board" of dilutions vs. incubation period, would take another week. Preparing your reagents can be done simultaneously with any others you have to prepare daily.
Finally you will have to analyze your results, select the best dilution, and make some "final adjustments"; that would be a third week.
So 3 weeks if that is the only thing you are doing (about 6h x 5 days x 3 weeks = 90 hours), BUT if you work less than 6 h/day in the project, extend the time to get to your 90 hours.
That is how I see the time neded for this project!
René J.
louise renton wrote:
Dear Histonetters,
First off, let me say what a great experience the NSH symposium was! It was
great to finally meet y'all. Made me realise that I am not the only one
suffering!
So here's my question rearding IHC.......
What is a reasonable work-up time for a new antibody? The parameters are:
1. Ab researh only, not used in the particular species of animal I am using
2. Ab titre needs to be assessed
3. not sure what a good control might be
4.Will be using archival tissue where fixation/processing may be
questionable
5. using a manual ABC/DAB technique
6. All buffers etc made up in-house
Any ideas??
Best regards
--
Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
"There are nights when the wolves are silent and only the moon howls".
George Carlin
No trees were killed in the sending of this message.
However, many electrons were terribly inconvenienced.
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