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. 2021 Mar 31;13(4):254. doi: 10.3390/toxins13040254

Figure 1.

Figure 1

Neutralization strategy for the characterization of anti-disintegrin against disintegrins. (A) Schematic representation of approach to characterize and neutralize disintegrins with anti-disintegrin. (B) Isolation of disintegrin D1. A total of 200 µL of venom solution (30 mg/mL) was injected into a Reverse Phase C18 column as described in the Materials and Methods. (C) SDS-PAGE analysis of the D1 peak collected by reverse phase HPLC. A total of 3 µg of D1 was run on 4–12% Bis-Tris Gel under non-reducing conditions at 100 V for 95 min. The gel was stained with SimplyBlue Safestain. lanes 1: SeeBlue Plus2 Markers, lane 2: D1. (D) N-terminal sequence of D1. The identity of the primary sequence was determined using Basic Local Alignment Search Tool (BLAST http://blast.ncbi.nlm.nih.gov/Blast.cgi (accessed on 29 March 2021)). (E) Amino acid sequence of C. atrox, C. horridus, and C. s. scutulatus disintegrins. Intra molecular cysteine linkages are proposed from Juarez et al. 2008 [18].