*2.2. Novel Toxins Identified by the Mass Spectrometry*

In order to identify the major proteins of *C. ihering*'s venom, their approximate molecular masses, and relative abundances, the crude venom was submitted for an SDS-PAGE, followed by a mass spectrometry analysis of the selected gel bands. The results revealed bands with apparent molecular masses ranging from 15 to 200 kDa, with proteins of the high molecular mass showing a higher intensity (Figure 3).

**Figure 3.** SDS-PAGE of the crude venom extracted from *C. iheringi*. Venom proteins were separated on a 12.5% SDS-PAGE gel and stained with Coomassie Brilliant Blue. The molecular mass (kDa) marker is shown on the right. The Selected groups of bands (from 1 to 6, on the left) were excised from the gel and processed for LC/MS-MS analysis.

Six groups of protein bands were selected and removed from the gel to be analyzed. The proteins were identified by matching the resulting mass spectra peptides to the deduced molecular masses of tryptic peptides, derived from full-length proteins as predicted from the transcriptome assembly. Matches of at least three peptides were considered valid. To identify a robust set of abundant venom toxins we focused on those sequences that exceeded the threshold value of 100 TPM. This allowed us to generate a list of 11 putative venom toxins that are highly abundant and likely play an important role in the venom (Table 3 and supplementary material Excel Table S1).


**Table 3.** Toxin identification of the major bands of *C. iheringi* venom based on the transcriptome and proteomic data. The numerical identifications correspond to the group of bands where the protein was found.


**Table 3.** *Cont.*
