Agarose gel matrix

Gel It usually uses an agarose gel matrix with alternating current in that the... 

Both the ease of use of this method for characterization of proteins and nucleic acids, and the abiHty to analyze many samples simultaneously for comparative purposes, have led to the prevalence of this technique. The drawbacks of a polyacrylamide matrix is that acrylamide is a neurotoxin, the reagents must be combined extremely carefiiUy, and the gels are not as pHable as most agarose gels. 

The double-immunodiffusion technique, often referred to as the Ouchtedony technique, uses an agarose gel as the matrix. Holes are made in the agarose where either sample or antisera is placed. The two solutions are allowed to diffuse into the matrix for a predetermined time. If there is a reaction... 

The affinities of antibodies for their cognate peptides can be very high, so that it is sometimes difficult to quantitatively recover the higher-affinity antibodies in a polyclonal serum from the peptide-matrix. For this reason, it is important to use low concentrations of peptide on the matrix (typically 100-200 pg of peptide per milliliter of agarose gel). In addition, elution of bound antibody in the reverse direction to which it was run into the column, so as not to drive eluting antibodies into a further excess of antigen, enhances recovery. If both of these criteria are adhered to, recovery is usually of the order of 60-80% see Note 3). 

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