Antibody out of stock: ab8580 - (Nov/25/2008 )

Nightmare....

I'm half way through the whole project and running low on this ab8580, Rb polyclonal against mouse trimethylated histone H3K4

And when I tried to order more, Abcam told me it's out of stock and could take up to YEARS to make more

they offer me ab1012 as an alternative....


But the thing is....should I redo everything with ab1012 now???

And does ab1012 work anyway?

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Sorry to hear about it. I've talked to a number of other people with the same problem. My guess is that a mouse monoclonal isn't going to work that great for ChIP (a few of the monoclonal antibodies we've used for ChIP work great but most don't) unless they certify it for ChIP (at least then you're covered if it doesn't work).

As far as redoing everything, I don't know how extensive your ChIP work is but that sounds like a lot of work. If you finish up with a different antibody rather than redoing everything you might find a way to normalize the signals, like expressing as fold enrichment over a negative control primer. Sorry again.

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For whoever might be interested:

ab1012 is about 15-fold less potent than ab8580....

I am now waiting for abcam to make me a new batch of ab8580

let's forget about ab1012

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Well, I don't know what region you're using the ab8580 to look at, but I used that antibody before on mouse cells, and the variation from lot to lot was HORRIBLE. The second lot I got from them was awful and didn't give me any signal at all. They did refund me the money... but... I really could have used a couple more experiments using that first lot, which they no longer have. So just keep in mind that polyclonals, although good for ChIP because of their binding ability, also has the variation problem. If you have some of the old lot left, make sure you keep some to do a parallel comparison ChIP to compare the two lots. That way you'll know that if your experiment fails with your new lot, it's not your technique, but the antibody... wish someone had told me that... =(

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FYI, we recently used an antibody from Diagenode (pAB-MEHAHS-024) for H4K4me3 that worked spectacularly well with our 96 well plate based ChIP protocol. I imagine it would likely work well for regular ChIP as well if you want to give it a try.

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