Journal: ACS Applied Materials & Interfaces
Article Title: Highly Specific Protein Identification by Immunoprecipitation–Mass Spectrometry Using Antifouling Microbeads
doi: 10.1021/acsami.1c22734
Figure Lengend Snippet: Coupling efficiency and antifouling performance of TA99 antibody to p SB- co -(azido) m % (pSB) beads. Panels B-G display flow cytometry data. (A) Evaluation of BCN-NHS coupling to TA99 antibody by SDS-PAGE analysis. Untreated TA99 antibody, TA99 labeled with BCN-NHS (BCN-TA99), and BCN-TA99 reacted with Azide-Lissamine (Liss-TA99) were visualized by fluorescence and Coomassie blue staining. (B) p SB- co -(TA99) m % beads with m = 2, 4, and 16%, TA99 attachment is shown using an anti-mouse-PE antibody. (C) p SB- co -(TA99) 8% beads in PBS are shown together with p SB- co -(TA99) 8% beads prepared using 0.5, 1, 2, 4, or 8 mg/mL TA99, TA99 was stained with an anti-mouse-PE antibody. (D) p SB- co -(azido) 8% (= 0% beads) and nonmodified (NM) beads in PBS, and NM and p SB- co -(TA99) m % ( m = 0, 2, 4, 16) beads incubated with serum-biotin followed by Strep-FITC. p SB- co -(TA99) m % beads were prepared using a 4 mg/mL TA99 antibody concentration. (E) p SB- co -(azido) 8% (pSB) in PBS, and p SB- co -(azido) 8% and p SB- co -(TA99) 8% (pSB-TA99) beads stained with an anti-mouse-PE (aMouse) antibody. (F) p SB- co -(TA99) 8% beads in PBS and incubated with BSA-AF488, as well as pSB and nonmodified (NM) beads incubated with BSA-AF488. Broad histogram peaks are most likely a result of inhomogeneous sample mixing, see the .
Article Snippet: Monoclonal TA99 antibody (IgG 2a ) was produced by a hybridoma cell line obtained from the American Tissue Culture Collection (ATCC HB-8704) that was cultured in Roswell Park Memorial Institute (RPMI) medium (Lonza), supplemented with l -glutamine, 10% fetal calf serum (FCS), 100 μg/mL penicillin, and 100 μg/mL streptomycin, at 37 °C and 5% CO 2 .
Techniques: Flow Cytometry, SDS Page, Labeling, Fluorescence, Staining, Incubation, Concentration Assay
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