US 8,262,887 B2Granted

Method for increasing measurement precision of two-dimensional protein

OwnerHan-Min Cheninventor

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Abstract

A method for increasing measurement precision of two-dimensional protein electrophoresis is provided, in which the electrical conductivity of a protein sample under test is measured for calculating the electrical energy required to enable salt and protein focusing. The method is characterized by a set of equations for calculating the electrical energy required respectively for protein focusing and for electrophoresis of salts in the protein sample, wherein the calculation is based on the electrical conductivity of the salts, the protein weight, a pH-gradient gel strip length, and a pH range. Thus, different protein samples can be supplied with the appropriate amounts of electrical energy for isoelectric focusing, so as to produce the optimal protein focusing effects and ensure that the focusing of protein in a gel will not be adversely affected by an otherwise insufficient or excessive supply of electrical energy.

A method for increasing measurement precision of two-dimensional protein electrophoresis, essentially comprising steps of:

(a) measuring a weight Mexp of a protein sample;
(b) measuring an electrical conductivity Sexp of salts in the protein sample;
(c) providing a set of equations for calculating a total electrical energy to be supplied to the protein sample, the set of equations including:
Vh
protein_exp
=
Vh
protein_ref
×
M
exp
M
ref
×
strip
⁢
⁢
length
exp
strip
⁢
⁢
length
ref
×
pH
⁢
⁢
range
ref
pH
⁢
⁢
range
exp
;
(
c1
)
Vh
salt_exp
=
Vh
salt_ref
×
S
exp
-
S
blank
S
ref
-
S
blank
×
strip
⁢
⁢
length
exp
strip
⁢
⁢
length
ref
×
pH
⁢
⁢
range
ref
pH
⁢
⁢
range
exp
;
(
c
⁢
⁢
2
)
⁢
and
Vhtotal=Vhprotein+Vhsalt;
(c3)
Vhtotal—exp=Vhprotein—exp+Vhsalt—exp (c4)
where Vhprotein—exp is an electrical energy for enabling protein focusing, to be determined by the equation (c1);
Vhsalt—exp is an electrical energy for enabling salt electrophoresis, to be determined by the equation (c2);
Vhprotein—ref is an electrical energy for enabling focusing of a reference protein;
Vhsalt—ref is an electrical energy for enabling electrophoresis of a reference salt;
Mref and Mexp are a weight of a reference protein and the weight of the protein sample, respectively;
Sblank, Sref, and Sexp are an electrical conductivity of a gel solution containing no protein, an electrical conductivity of the reference salt, and the electrical conductivity of the salts in the protein sample, respectively;
strip lengthref and strip lengthexp are a reference pH-gradient gel strip length and an actual pH-gradient gel strip length, respectively;
pH rangeref and pH rangeexp are a reference pH range and an actual pH range, respectively;
and Vhtotal, Vhprotein, and Vhsalt are the total electrical energy, an electrical energy for enabling protein focusing, and an electrical energy for enabling salt electrophoresis, respectively;
and
(d) substituting the electrical conductivity of the salts in the protein sample, the weight of the protein sample, the actual pH-gradient gel strip length, and the actual pH range into the set of equations so as to determine the total electrical energy for enabling protein focusing and achieving optimal protein focusing effects.

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Abstract

Claims7