Proteins 350 and 780 were constructed and expressed as previously reported (Gray et al. 2012 (link)). Proteins 030 and 14-3-3 were obtained from the Seattle Structural Genomics Center for Infectious Disease (SSGCID). All four antigens were biotinylated using either the Pierce EZ-Link NHS-Peg 4–Biotin or the Sulfo-NHS-LC-Biotin kit (Thermo Scientific, Rockford, IL). Biotinylation was confirmed by Western blots probed with 0.4 ug/mL HRP-conjugated streptavidin (Sigma Aldrich, St. Louis, MO).
Sulfo nhs lc biotin kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Sulfo-NHS-LC-Biotin kit is a reagent used for biotinylation of proteins. It contains sulfo-NHS-LC-biotin, a water-soluble, amine-reactive biotin derivative that can be used to label primary amines on proteins.
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Lab products found in correlation
2 protocols using sulfo nhs lc biotin kit
1
Biotinylation of Protein Antigens
2
CAR T-cell surface expression analysis
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On day 0, CEA-CD28-CD3z CAR transgenic CD3 + T cells (10 7 cells) engineered with or without IL-18 were injected subcutaneously (s.c. For detection of CAR expression on the surface of Tregs, sections were incubated with the hybridoma-derived anti-idiotypic antibody BW2064/36 previously biotinylated by the Sulfo-NHS-LC-Biotin kit (Thermo Fisher Scientific, Waltham, MA, USA). Following incubation with the primary antibody, antibody binding was visualized using AF-647-labeled streptavidin (BioLegend). Slides were analyzed using an Olympus FV 1000 microscope (Olympus, Tokyo, Japan), and the fluorescence intensity was determined using ImageJ software.
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