Hi
I saw the interesting discussion of dissolving PP1 in PBS.
I always take it for granted that one can dissolve water insoluble stuff in an organic solvent then add it to a water based solution. But I have never thought about it. Why is that possible? Why it wont precipitate out from the solution? That should still depends on the compound's solubility in water. Does it mean the organic solution is simply for a high concentration stock?
BTW, is there a database for drug solubility?
have a nice Xmas
SZ
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Subject: Methods Digest, Vol 43, Issue 16
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Today's Topics:
1. Re: An inquiry (Dr. Hiranya S. Roychowdhury)
2. RNAi (Jayanta Tarafdar)
3. methylcellulose colony question (Scott Brown)
4. RACE sequencing problem (Scott Brown)
----------------------------------------------------------------------
Message: 1
Date: Fri, 19 Dec 2008 10:29:33 -0700 (MST)
From: "Dr. Hiranya S. Roychowdhury" <hroychow from nmsu.edu>
Subject: Re: An inquiry
To: "Shahrzad Jalali" <shahrzadjalali from gmail.com>
Cc: methods from magpie.bio.indiana.edu
Message-ID: <1240.71.210.225.55.1229707773.squirrel from webmail.nmsu.edu>
Content-Type: text/plain;charset=iso-8859-1
I don't believe you can expect a solution of PP1 in PBS. It will form a
colloidal suspension. A colloidal suspension will also elicit immune
response. Remember that the peritonial fluid is also polar and the PP1
will come out of solution there too. DMSO, however, does the rat very
little damage in microliter quantities intra-peritonially. I believe the
PBS is used to minimize any shock. PP1 is soluble also in ethanol, and it
may be a better alternative solvent for the injection. Just a thought!
> To whom it may concern,
>> I am from U of Toronto and have recently joined to your group. Had some
> problems in dissolving a compound and was wondering if I can get any
> suggestion from the expert people.
>> I have to do i.p. injections to the rats and the compound of interest is
> src
> kinase inhibitor PP1.
>http://www.biomol.com/SiteData/docs/productdata/ei275.pdf. As I got the
> information from the literature, it has been mentioned in several papers
> that this material is dissolved in DMSO and further diluted in the PBS
> prior
> to the injection to rats.
> In the data sheet of the company it is also mentioned that this material
> should be dissolved in DMSO (25 mg/ml). Each vial had 5mg in it, I
> initially added 200ul of DMSO to that and completely dissolve it by
> pipetting up and down, but when I added PBS (as indicated in the papers),
> all the compound was precipitated and I got just a colloidal solution. The
> company did not have any suggestion, as they told that it is hydrophobic
> material. I don't know how the papers have done that. On the other hand I
> have to dilute it in PBS because injecting DMSO to the rats has other
> effects which can interfere with my experimental plan. I should mention
> that
> heating the compound up to 37C for several hours and its repetitive
> agitation did not help this compound go to the solution.
>> I would appreciate if anybody can give suggestion in this regard and
> urgently waiting for the nswer,
> Thanks,
> Shahrzad
> _______________________________________________
> Methods mailing list
>Methods from net.bio.net>http://www.bio.net/biomail/listinfo/methods>
--
Hiranya S. Roychowdhury, Ph.D.
Asst. Professor,
Health & Public Services
Dona Ana Community College
New Mexico State University
Las Cruces, NM 88003
------------------------------
Message: 2
Date: Fri, 19 Dec 2008 23:23:03 +0530
From: "Jayanta Tarafdar" <jayanta.tcbckv from gmail.com>
Subject: RNAi
To: methods from magpie.bio.indiana.edu
Message-ID:
<be8bf3be0812190953i3c458dfm296893a5f4357ec0 from mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Dear Subscribers
Methods Digest
We have been working on the backcross breeding of rice with transgenic
rice
carrying RNAi gene against virus. Would anybody help me regarding use of
non-radioactive probe for analysis of the transcript.
I would appreciate if anybody can give short-cut method in this regard
and
urgently waiting for the answer.
Jayanta
India
------------------------------
Message: 3
Date: Sat, 20 Dec 2008 04:33:48 +1100
From: "Scott Brown" <SBrown from ccia.unsw.edu.au>
Subject: methylcellulose colony question
To: <methods from magpie.bio.indiana.edu>
Message-ID:
<2A67EA781EC7F949A2AB0A0D07A86C6A02972BC5 from mail01.ccia.local>
Content-Type: text/plain; charset="iso-8859-1"
Hi,
I have been growing Nalm-6 colonies in methocult for about a year now, the ratio of media is 1.6ml methocult, 1.2ml IMDM and 1.2ml FCS with or without drug depending on what i am doing.
Has anyone had any experience with maintaining the methocult for at least 3 weeks, frequently my methocult looks as though is has dessicated, i have water in the incubator and i also plate 2 wells with PBS to ensure humidity...but...it still happens, any ideas or suggestions?
S
Scott Brown
PhD Candidate
Children's Cancer Institute Australia for Medical Research is the only independent medical research institute in Australia devoted to research into the causes, prevention, better treatment and ultimately a cure of childhood cancer. Our vision is to save the lives of all children with cancer and to eliminate their suffering.
The information contained in this message and any annexure is confidential and intended only for the named recipient(s). If you have received this message in error please contact the sender immediately and destroy the original message.
------------------------------
Message: 4
Date: Sat, 20 Dec 2008 04:38:40 +1100
From: "Scott Brown" <SBrown from ccia.unsw.edu.au>
Subject: RACE sequencing problem
To: <methods from magpie.bio.indiana.edu>
Message-ID:
<2A67EA781EC7F949A2AB0A0D07A86C6A02972BC6 from mail01.ccia.local>
Content-Type: text/plain; charset="iso-8859-1"
Hi,
I have been sequencing constructs about 1kb in size that i have obtained from a second race reaction, i have pruified the product, spec'd it and ensured when run on a gel i get a band.
This has worked for a while, however the last 1 or so constructs i have had sequenced have failed, the chromatogram looks as though the primer has not even bound to the template, it is simply noise that drops off to an almost undetectable signal.
Can anyone make any suggestions as to what I should be looking at to troubleshoot this? I sent my positive control as well as 1 sample from my next set of rave reactions to be sequenced, both were successful, then when i sent the remaining 8, all 8 failed.
Might i need to redesign my primer i am using for sequencing? Any other thoughts?
S
Scott Brown
PhD Candidate
Children's Cancer Institute Australia for Medical Research is the only independent medical research institute in Australia devoted to research into the causes, prevention, better treatment and ultimately a cure of childhood cancer. Our vision is to save the lives of all children with cancer and to eliminate their suffering.
The information contained in this message and any annexure is confidential and intended only for the named recipient(s). If you have received this message in error please contact the sender immediately and destroy the original message.
------------------------------
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