Re: 2D : question concerning abnormalities in 2D-electrophoresis
Dirk S. Krapf (krapf@abrf.org)
Thu, 13 May 1999 08:40:01 -0700
>---------- Forwarded message ----------
>Date: Wed, 12 May 1999 16:16:30 +0200
>From: Jos Raymackers <Jos_Raymackers@innogenetics.com>
>To: Recipients of ABRF List <abrf@aecom.yu.edu>
>Subject: 2D : question concerning abnormalities in 2D-electrophoresis
>
>
>Dear all,
>
>I have some questions for you concerning 2D-electrophoresis. Here they come
>:
>
>1.
>Have you ever seen the phenomenon that your IPG-strip has a "bubble" in the
>middle of the strip (approximately in the middle). It seems that the gel
>loosened from the plastic and/or that there are some airbubbles between the
>gel and the plastic. It appears during the first dimension run (on
>Multiphor) and it 's still there after the first separation is done. Has it
>something to do with the saltconcentration ? In this case it was 9.6 mM
>(>10mM !!). Or has it something to do with electro-endo-osmosis ( I blotted
>the electrodewicks before, so they were not too wet !). I must mention
>that I used 7 cm long IPG-strips in this case but I have seen it also in 18
>cm long strips. Do you know the reason for this phenomenon and do you know
>how to avoid it ?
>2.
>Did you ever mentioned that when using the IPG-strips 4-7 for 2D, the spots
>in the acidic area were rather diffuse ? What may be the reason for that ?
>3.
>I also attached some tif.files of two 2D-patterns. They were run in a batch
>of 10 gels. Some of these gels were OK, but some of them had some artefact
>(see images; I see some vertical streaking (a bit flam-shaped (arrow)),
>spots become diffuse, but first dimensional separation seems to be
>succeeded (see spots on top of image zipad04j)). It's becoming a big
>problem, since I have no idea what causes it, and it also appears in other
>batches. It 's a bit strange because some of the gels run in one batch are
>good and some of them are not, so it must be caused when treating the
>strips separately (e.g. the rehydration or the equilibration), I think. I
>thought about detergent left after cleaning the rehydration/equilibration
>trays ??? I use the Multiphor and the ISO-Dalt as equipment. Do you
>recognise the artefact and/or do you have a suggestion to solve this
>problem ?
>
>(See attached file: Zipad04e.tif)(See attached file: Zipad04j.tif)
>
>Thanks in advance !!!
>Answers may be send to Jos Raymaekers or to annicdan@innogenetics.be
>
>Kind regards,
>
>Annick Dani?ls,
>Innogenetics N.V.
>Industriepark Zwijnaarde 7, box 4
>B-9052 Gent
>Belgium
>
>Phone : 0032-9-241.07.77. or 78
>e-mail : annicdan@innogenetics.be
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