How to make the starch gel for STARCH GEL ELECTROPHORESIS?  I have
solubel starch with specifications matching to the specifications of
starch for gel electrophoresis, given in Sigma Aldrich Catalog. How do
i prepare the gel for starch gel electrophoresis? What is the solvent
that i use? Will water be fine as solvent? What concentration will be
required?

I have never done ?Starch Gel Electrophoresis, so I can?t advise you
on the technical aspects of this but I have worked with starch before
so I can tell you how to mix it. First get a container at least 50%
greater volume than the liquid you will need and add the correct
volume of solvent (water usually, which may need to be deionised for
your purposes). You are going to need a powerful mixer, one that looks
a bit like a kitchen blender but much bigger and it must have a
bearing at both the top and bottom of the shaft. The shaft naturally
wants to whip so the bottom bearing, (the one in the liquid) tends to
go first so check that this good before you start. The reason you will
need a powerful motor and good bearings is that the motor will want to
maintain its speed but the liquid will become more and more viscous so
there is more strain put on the system. Set the mixer so the shaft is
in the solvent container and the impeller just clears the bottom and
switch on. A vortex will be created and the solvent will rise up the
sides of the container which is why it needs to be at least 50%
bigger. Throw the starch as quickly as you like into the solvent BUT
NOT INTO THE VORTEX otherwise it will clag. Keep mixing for about
twenty minutes and your solution will be ready.

I am sure you are aware that as well as normal neutral starch, you can
also obtain both cationic and anionic modified starches which may be
more useful for your purpose.

Other than this, the large starch manufacturers have technical service
departments that should be able to help you. There is not much that is
100% new so they should be to come up with some good advice. Even if
you are not buying from them now you could do in the future!

Hope this is at least a little helpful.

cf

Starch gels are usually used to look at isozyme patterns. This means
you need a buffer that will keep the molecules in their native form.
This allows them to be separated by variations in charge, folding
etc., and remain functional, so you can test for activity in the gel.
Because the conditions will change, depending on which enzyme you want
to look at, there is no single buffer that is universal. The standard
work for all this is Shaw, C.R., and R. Prasad, 1970. Starch gel
electrophoresis of enzymes - A compilation of recipes. Biochemical
Genetics 4:297-320
To give you some idea of the amount of variation here are some buffers
and running conditions for some enzymes. For each the information is
given in order
Electrode buffer
Gel buffer
Running conditions
Enzyme system

I
TRIS-Citrate system  
0.148 M Tris  
0.047 M citric acid  
pH 7.3
11-12% concentration.  
0.038 M Tris  
0.012 M citric acid
approx. 20 V cm-1 for 4 h, max. 180 mA or max. 200 Volt.
PGI, PGM, IDH, MNR, SKDH, MDH, 6-PGDH, ADH.

II
ASHTON-system  
0.19 M boric acid  
0.042 M lithium hidroxide  
pH 8.1
11-12 % concentration  
0.05 M Tris  
9.5 mM citric acid  
pH 8.1
approx. 18 V cm-1 for 5 to 5 h, max. 200 Volt. or 80 mA.
AAT  
LAP  
MNR  
PGM

III
POULIK-system  
0.3 M Tris  
NaOH 10N  
pH 8.0
9-10% concentration  
0.017 M Tris  
2.3 mM citric acid  
pH 8.0
approx. 18-20 V cm-1 for 5 h, max. 200 Volt. or 70 mA
AAT  
LAP  
MNR  
PGM

IV
TRIS-Histidine-system  
0.048 M L-Histidine HCL  
9.5 mM citric acid  
pH 6.5
9-10% concentration  
7.2 mM L-Histidine HCL  
1.4 mM citric acid  
pH 6.5
approx. 18-20 V cm-1 for 4 h, 15-30 mA or max. 200 Volt.
PGI, PGM, IDH, MNR, SKDH, MDH, 6-PGDH, ADH.