Serum (antigen) when heated at a temperature sufficient to cause definite clouding reacts more intensely with a specific precipitin than a portion of the unheated serum or samples heated at lower temperatures. The phenomenon is explained on the basis that coagulated protein in suspension is covered with undenatured antigen and the addition of precipitin causes agglutination of the coagulated protein. Similar phenomena are obtained when bacteria or collodion particles are mixed with diluted serum (antigen) and precipitin added; the particles or bacteria agglutinate and increase the visibility of the reaction.
Further, it is shown that collodion particles sensitized with cow serum or crystallized egg albumin and subsequently washed until the washing fluid no longer contains the antigenic substance will agglutinate when small quantities of specific precipitin are added. Bacteria sensitized with cow serum and subsequently washed until cow serum no longer remains in the washing solution agglutinate when cow antiserum at fairly low concentration is added. It was not possible to show that bacteria soaked in crystallized egg albumin and subsequently washed retained on their surfaces sufficient undenatured egg albumin to react to crystallized egg albumin precipitin.