Help solubilize insoluble protein

Akihiro Ohno, Ph. D.

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Jan 30, 1997, 3:00:00 AM1/30/97

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Dear fellow scientists:
I am currently in need of protocol to solubilize protein which is
insoluble to water. It was once soluble to water, but after ammonium
sulfate precipitation followed by washing with deionized water on the
ultrafilteration membrane (50000cut), it has become insoluble.
For our next experimental step, we have to inject the desired protein
to rats, so we cannot use toxic detergents nor organic solvents for
solubilization.
Could you suggest some methods/reagents which might enable
solubilization of the protein while keeping it non-toxic to the animals?
Thank you for your suggestion.
--
+++++++++++++++++++++++++++++++++++++++++++++++++++++++
Akihiro Ohno, Ph. D.
Advance, Co., Ltd., Tokyo, Japan
E-mail: ohn...@bnn-net.or.jp

nes...@earthlink.net

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Jan 30, 1997, 3:00:00 AM1/30/97

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"Akihiro Ohno, Ph. D." <ohn...@bnn-net.or.jp> wrote:

>Dear fellow scientists:
> I am currently in need of protocol to solubilize protein which is
>insoluble to water. It was once soluble to water, but after ammonium
>sulfate precipitation followed by washing with deionized water on the
>ultrafilteration membrane (50000cut), it has become insoluble.
> For our next experimental step, we have to inject the desired protein
>to rats, so we cannot use toxic detergents nor organic solvents for
>solubilization.
> Could you suggest some methods/reagents which might enable
>solubilization of the protein while keeping it non-toxic to the animals?
>Thank you for your suggestion.

Akihiro Ohno:

Formic acid is one of the best reagents for solubilizing peptides and
proteins. Use a small volume of 30-50% formic acid to solubilize your
pellet. Then dilute the formic acid with the buffer of your choice
for you in vivo studies. Hope this helps.

Best regards,

John

**********************************
John K. Troyer, Ph.D.
The Nest Group, Inc.
nes...@earthlink.net
http://world.std.com/~nestgrp
**********************************

Ralf Spenneberg

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Jan 31, 1997, 3:00:00 AM1/31/97

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In article <32F004...@bnn-net.or.jp>, "Akihiro Ohno, Ph. D."
<ohn...@bnn-net.or.jp> wrote:

> Dear fellow scientists:
> I am currently in need of protocol to solubilize protein which is
> insoluble to water. It was once soluble to water, but after ammonium
> sulfate precipitation followed by washing with deionized water on the
> ultrafilteration membrane (50000cut), it has become insoluble.
> For our next experimental step, we have to inject the desired protein
> to rats, so we cannot use toxic detergents nor organic solvents for
> solubilization.
> Could you suggest some methods/reagents which might enable
> solubilization of the protein while keeping it non-toxic to the animals?
> Thank you for your suggestion.

> --
Hi!
Just in case you want to induce antibodies: Why can't you use the
insoluble Protein. Usually insoluble proteins make very good antigens.
If that's not want you are trying to do, what exactly is it? Where do you
inject the rats (i.p., s.c., i.m. i.v.)?
One thing you might try is to disolve the protein in 8M Urea and to
dialyse the Urea.

Ralf.

--
Ralf Spenneberg
Institute for Medical Biochemistry
Universitaet Muenster 0251 83 6722
Von Esmarchstr 56 48149 Muenster Germany