Nonfat Milk Powder & Western Blot
Stefan Beyreuther
Hi Bionet Folks.
I'd like to use milk powder in a western blot instead of bsa (bovine
serum albumine) for the blocking of the membrane.
As far as I know nonfat milk powder has to be used for that purpose.
Here in Germany I could get low fat powder that still contains 1 % fat.
So the question is: What is nonfat milk powder? Does it contain zero %
fat? Or does it just mean to contain only 1 % so that the stuff I got
here would be ok?
Does anybody know? Or at least have an idea where I could get a hint if
not here?
Best regards
Stefan
Stefan Beyreuther
Inorganic Chemistry Department
University of Heidelberg
Germany
Keld Sorensen
Jest remember that milk contains varying amounts
of biotin and phosphoproteins, so if that is part of
your blot (avidin/biotin or phosphoprotein detection)
you should not use milk.
Also it is highly recommended to try a couple of different
blocking reagents - blocking is not as simple as some
researchers think - every so often you run into
unexplained differences - each system is different.
Dr E. Buxbaum
Stefan Beyreuther <ste...@indi.aci.uni-heidelberg.de> wrote:
>Hi Bionet Folks.
>
>I'd like to use milk powder in a western blot instead of bsa (bovine
>serum albumine) for the blocking of the membrane.
>
>As far as I know nonfat milk powder has to be used for that purpose.
>Here in Germany I could get low fat powder that still contains 1 % fat.
Go for it. Never had any problems with that stuff.
Anonymous
mmmm
In article <jnakamot-181...@news.ucla.edu>, jnak...@ucla.edu
(Jon Nakamoto) wrote:
> Whatever you decide to use, I'd run a comparison of two or three milk
> powders beforehand. I remember how variable results could be (Carnation
> looked more yellowish than the others and gave horrible results; Sanalac
> and Ralphs market's generic brand worked great).
>
> Jon
>
> Jon Nakamoto, MD
> Asst Prof of Pediatrics/Endocrinology
> UCLA Children's Hospital
> jnak...@ucla.edu
>
>
>
>
> In article <32879D...@indi.aci.uni-heidelberg.de>,
> ste...@indi.aci.uni-heidelberg.de wrote:
>
> > Hi Bionet Folks.
> >
> > I'd like to use milk powder in a western blot instead of bsa (bovine
> > serum albumine) for the blocking of the membrane.
> >
> > As far as I know nonfat milk powder has to be used for that purpose.
> > Here in Germany I could get low fat powder that still contains 1 % fat.
> >
> > So the question is: What is nonfat milk powder? Does it contain zero %
> > fat? Or does it just mean to contain only 1 % so that the stuff I got
> > here would be ok?
> >
> > Does anybody know? Or at least have an idea where I could get a hint if
> > not here?
> >
> > Best regards
> > Stefan
> >
> > Stefan Beyreuther
> > Inorganic Chemistry Department
> > University of Heidelberg
> > Germany
>
> --
> Jon Nakamoto
> jnak...@ucla.edu
Jon Nakamoto
Alix Helme-Guizon
On Mon, 18 Nov 1996, Jon Nakamoto wrote:
> Whatever you decide to use, I'd run a comparison of two or three milk
> powders beforehand.
Any commercial so called "non fat" milk could be used. just avoid to use
any milk with additionnal vitamins or ions (especially calcium!).
I use 0.2% lipid containing milk in variable concentration and providers
without trouble except with calcium added milk.
Best regards,
Alix.
Toumy Guettouche
On Mon, 18 Nov 1996, Jon Nakamoto wrote:
> Whatever you decide to use, I'd run a comparison of two or three milk
> powders beforehand. I remember how variable results could be (Carnation
> looked more yellowish than the others and gave horrible results; Sanalac
> and Ralphs market's generic brand worked great).
>
> Jon
>
> Jon Nakamoto, MD
> Asst Prof of Pediatrics/Endocrinology
> UCLA Children's Hospital
Well, talking about variable results. In my comparison, carnation was the
best one (compared to Publix brand and two others I can not remember but
they were regular brands).
Toumy
Robin Colgrove
Alix Helme-Guizon wrote:
>
> On Mon, 18 Nov 1996, Jon Nakamoto wrote:
>
> > Whatever you decide to use, I'd run a comparison of two or three milk
> > powders beforehand.
> any milk with additionnal vitamins or ions (especially calcium!).
> I use 0.2% lipid containing milk in variable concentration and providers
> without trouble except with calcium added milk.
> Best regards,
> Alix.
Actually, I always used carnation non-fat dry milk with vitaimins A&D
to good effect and it was a lab joke to cite NFDMAD as the key blocking
reagent with that magical extra little kick from the carotenoids ;^)
btw, does anybody else remember that classic biotechniques article
from the late '80s describing how to block with Baileys Irish Cream?
The author, some big name I think but I am "blocking" on who, describes
how
the leftovers can be used for either celebration or commiseration
depending on the results of the experiment.
Hmmm, now that I think about it, maybe it was for blocking southern's
rather than westerns. Must be too much Baileys in my own experiments...
robin
Paul N Hengen
Robin Colgrove (ro...@louis.bidmc.harvard.edu) wrote:
> btw, does anybody else remember that classic biotechniques article from the
> late '80s describing how to block with Baileys Irish Cream? The author, some
> big name I think but I am "blocking" on who, describes how the leftovers can be
> used for either celebration or commiseration depending on the results of the
> experiment. Hmmm, now that I think about it, maybe it was for blocking
> southern's rather than westerns. Must be too much Baileys in my own
> experiments...
I knew about the Blotto technique, but the Baileys Irish Cream is a new
one on me. Oh, please please somebody find me the reference for this one.
--
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* National Cancer Institute |Internet: p...@ncifcrf.gov |*
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Michael Benedik
In article <5758fh$k...@ncisun1-nf0.ncifcrf.gov>
I remember the article. It was from Bert O'Malley's lab at Baylor.
Don't have the reference and the Biotechniques search site is not
working now.
Michael Benedik
Department of Biochemical Sciences
University of Houston
ben...@uh.edu