Carbon nanotube (CNT)-loaded and neat polyacrylonitrile nanofibers were produced by a needleless continuous electrospinning method as carbon nanofiber precursors. The details of the stabilization, which is a crucial issue during carbon fiber production, were investigated as these nanofibers are especially sensitive to degradation. In order to determine the optimal parameters, the nanofibers were stabilized at different temperatures. The stabilized samples were analyzed by Fourier-transform infrared spectroscopic and differential scanning calorimetric (DSC) measurements and by the determination of the color changes. The chemical changes during the stabilization (the formation of the so-called ladder-polymer) can be followed by infrared spectrometry, while the conversion can be monitored by DSC. The formation of the ladder-polymer occurs according to the Gaussian distribution function, where the temperature of the stabilization is the statistical parameter, which was also determined. In the case of CNT-loaded samples, the range of stabilization temperature was wider, which provides better controllability of the process. Based on the established models, an appropriate multi-step heat-treatment program could be determined, which led to completely stabilized nanofibers, suitable for carbonization.