Journal: The Journal of Biological Chemistry
Article Title: Evaluation of AlphaFold modeling for elucidation of nanobody–peptide epitope interactions
doi: 10.1016/j.jbc.2025.110268
Figure Lengend Snippet: Evaluation of a crosslinking peptide for Nb 127 guided by modeling and structure. A, structure of an analog of the 127-peptide tag with a crosslinking group (X) incorporated. B, mass spectrometry analysis of Nb 127 before ( left ) and after ( right ) reaction with Ac-127X3. Crosslinking was performed through mixing Nb 127 (10 μM) and Ac-127X3 (20 μM) in PBS for 2 h at 25 °C followed by purification using a disposable size-exclusion column (PD10). C, assessment of the time-dependent formation of a covalently crosslinked complex from Nb 127 and FAM-127X3 via detection of a fluorescent band on SDS-PAGE. D, assessment of the reaction of FAM-127X3 and Nb 6E (negative control) as described in C . Fluorescent gel images were obtained as described in the section. E, quantification of formation of a fluorescent crosslinking product through densitometric analysis of experiments described for C and D . Fluorescence intensity was quantified using densitometry as described in the section. The data points are presented as mean ± SD of two independent experiments. An uncropped gel image for data in C and D , as well as corresponding Coomassie staining, is shown in . Nb, nanobody.
Article Snippet: The gel was first scanned to detect fluorescence signals, followed by total protein staining using Coomassie stain (Protein Gel Stain; Biotium, catalog no.: 21003).
Techniques: Mass Spectrometry, Purification, SDS Page, Negative Control, Fluorescence, Staining
Biotium is a verified supplier 
Biotium manufactures this product 
