Journal: Scientific Reports

Article Title: Anti-IgD nanobodies as novel tools for studying human IgD biology

doi: 10.1038/s41598-025-09118-4

Figure Lengend Snippet: Interaction of anti-IgD Nbs with IgD. Binding of anti-IgD Nbs ( a ) aδNb107, ( b ) aδNb367, ( c ) aδNb408 and ( d ) aδNb571 to IgD captured on an anti-His chip by SPR. A two-fold dilution series of TEV-cleaved anti-IgD Nbs was flowed over His-tagged IgD, from the highest concentration 200 nM (black line) to the lowest concentration 1.6 nM (purple line). Binding of IgD-Fc to anti-IgD Nbs ( e ) aδNb107, ( f ) aδNb367, ( g ) aδNb408 and ( h ) aδNb571. The His-tagged Nbs were captured by anti-His. A two-fold dilution series of IgD-Fc was flowed over, with the highest concentration 200 nM (black line) and the lowest concentration 3 nM (green line). Double-reference subtraction was not performed for ( e ) aδNb107, ( f ) aδNb367 and ( g ) aδNb408, with the 0 nM concentration shown as a gray line. RU, resonance units.

Article Snippet: Anti-IgD Nbs aδNb107 (Addgene 220319), aδNb367 (Addgene 220320), aδNb408 (Addgene 220321) and aδNb571 (Addgene 220322) in pET-15b were codon-optimized for E. coli expression and synthetized by GenScript, with an N-terminal periplasmic leader and a C-terminal TEV cleavage site, followed by a glycine-serine linker and His 6 -tag.

Techniques: Binding Assay, Concentration Assay

Journal: Scientific Reports

Article Title: Anti-IgD nanobodies as novel tools for studying human IgD biology

doi: 10.1038/s41598-025-09118-4

Figure Lengend Snippet: Anti-IgD Nbs as tools for SPR capture. Biotinylated aδNb107, aδNb367 and aδNb408 were immobilized onto an SA chip. ( a ) IgD-Fc was captured by anti-IgD Nbs and a two-fold dilution series of aδNb571 was flowed over, with the highest concentration 50 nM aδNb571 (orange line) and the lowest concentration 3 nM aδNb571 (green line). ( b ) Full-length IgD, HAPPID1, was captured by anti-IgD Nbs and a two-fold dilution series of the polcalcin allergen Ole e 3 was flowed over, with the highest concentration 100 nM (cyan line) and the lowest concentration 1.6 nM (purple line). RU, resonance units.

Article Snippet: Anti-IgD Nbs aδNb107 (Addgene 220319), aδNb367 (Addgene 220320), aδNb408 (Addgene 220321) and aδNb571 (Addgene 220322) in pET-15b were codon-optimized for E. coli expression and synthetized by GenScript, with an N-terminal periplasmic leader and a C-terminal TEV cleavage site, followed by a glycine-serine linker and His 6 -tag.

Techniques: Concentration Assay

Journal: Scientific Reports

Article Title: Anti-IgD nanobodies as novel tools for studying human IgD biology

doi: 10.1038/s41598-025-09118-4

Figure Lengend Snippet: Anti-IgD Nb pairs as a tool for cell-binding. ( a ) Left: Flow cytometry data of AF488-labeled anti-IgD Nbs binding to Namalwa cells. A non-IgD binding Nb was used as a control for non-specific background binding. Right: MFI was normalized to the control and the highest binder aδNb367, with the mean plotted as a gray bar and the individual data points as black crosses (n = 3). ( b ) Left: Flow cytometry data of anti-IgD Nb pairs binding to Namalwa cells. A pair of non-IgD binding Nbs was used as a control for non-specific background binding. Right: MFI was normalized to the control and the highest binder aδNb408-aδNb107, with the mean plotted as a gray bar and the individual data points as black crosses (n = 2). ( c ) Comparison of the bivalent/bispecific aδNb408-Nb constructs (panel b, left) with their monovalent Nb components (panel a, left); these data are from the same experiment and are therefore comparable. MFI values of monovalent aδNb107 (orange, 662), monovalent aδNb367 (blue, 1452), monovalent aδNb408 (magenta, 1339) and monovalent aδNb571 (green, 921) are shown additively as filled bars, separated by a dashed line. MFI values of equivalent bivalent/bispecific Nbs are shown as empty black bars: aδNb408-aδNb107 (3533), aδNb408-aδNb367 (2641), aδNb408-aδNb408 (1998) and aδNb408-aδNb571 (2488). ( d ) Flow cytometry data of Nb pair aδNb408-aδNb107 titrated on Namalwa cells. MFI, median fluorescence intensity; AF488, Alexa Fluor 488; A.U., arbitrary units.

Article Snippet: Anti-IgD Nbs aδNb107 (Addgene 220319), aδNb367 (Addgene 220320), aδNb408 (Addgene 220321) and aδNb571 (Addgene 220322) in pET-15b were codon-optimized for E. coli expression and synthetized by GenScript, with an N-terminal periplasmic leader and a C-terminal TEV cleavage site, followed by a glycine-serine linker and His 6 -tag.

Techniques: Binding Assay, Flow Cytometry, Labeling, Control, Comparison, Construct, Fluorescence

Journal: Molecular cell

Article Title: MRI is a DNA Damage Response Adaptor during Classical Non-Homologous End Joining

doi: 10.1016/j.molcel.2018.06.018

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: WT (lines SZ and WT-1), MRI −/− (lines M61.2 and M61.7), and XLF −/− (lines X-AB and X-SZ) MEFs were generated from E14.5 or 15.5 mice and immortalized by transfection with pBABE-neo-SV40 (Addgene), after which they were selected in 400 μg/mL G418 (Thermo Fisher).

Techniques: Blocking Assay, Recombinant, Cell Isolation, Magnetic Beads, Mass Spectrometry, Magnetic Resonance Imaging, Sequencing, Plasmid Preparation, Software, High Content Screening, Flow Cytometry, Inverted Microscopy, Laser-Scanning Microscopy, Spectrophotometry, Irradiation