monoclonal antibody  (R&D Systems)

Journal: Nature Communications

Article Title: Targeting mGlyR with nanobodies for depression

doi: 10.1038/s41467-026-68339-x

Figure Lengend Snippet: a Schematic of the nanobody development pipeline. A phage library was constructed from leukocytes of a llama immunized with mGlyR, followed by 3 rounds of panning on mGlyR to enrich for specific binders, which were isolated and tested. b Schematic of the detection strategy in flow cytometry experiments. c Analysis of nanobody binding by flow cytometry of HEK cells transiently expressing mGlyR incubated with or without Nb20 and anti-myc-APC conjugated antibody. Percentages of cells in each quadrant are indicated. d Dose-response profiles of representative Nb20 binding experiment to cells expressing mGlyR in flow cytometry experiments. Concentrations of Nb20 are shown. e Quantification of data in panel D. Error bars are SEM values ( n = 3 independent experiments). f Schematic of the surface plasmon resonance (SPR) assays that detect Nb20 binding to the chip containing immobilized recombinant ectodomain of mGlyR (Ecto-mGlyR). g SPR sensorgram of binding and dissociation of Nb20.

Article Snippet: Nanobody-20 (Nb20) and 10 μl of anti-myc-APC conjugated antibody (R&d Systems #IC3696A) were added and incubated in the dark with rotation, at 4 °C for 1 h. After 3 washes, cells were analyzed in the LSR-II BD flow cytometer.

Techniques: Construct, Isolation, Flow Cytometry, Binding Assay, Expressing, Incubation, SPR Assay, Recombinant

Journal: Scientific Reports

Article Title: A novel luciferase-based assay for the detection of Chimeric Antigen Receptors

doi: 10.1038/s41598-018-38258-z

Figure Lengend Snippet: ( A ) Non-transduced JNG cells and JNG cells stably expressing the indicated BCMA-CARs (Centyrin, J6MO, and BB-CAR-02) and a CD19-CAR (FMC63) were incubated with 100 µl of supernatant containing the BCMA-ECD-Nluc fusion protein for 45 minutes on ice. After incubation, cells were washed 5 times and assayed for luminescence. ( B ) The assay was performed as described in ( A ) except that CD19-ECD-Nluc fusion protein was used. ( C ) JNG cells stably expressing the indicated CARs were incubated with an APC-conjugated MYC antibody for 45 minutes on ice. After incubation, cells were washed twice and analyzed by flow cytometry. ( D ) JNG cells stably expressing the indicated CARs were stained with Protein L reagent as described in methods section followed by flow cytometry analysis. The values shown are mean ± SE of a representative of at least two experiments performed in duplicate.

Article Snippet: Strep-Tactin-APC (IBA-lifescience: Cat# 6-5010-001); Anti-Flag-FITC (Sigma: F4049); Anti-Strep-tagII-FITC (Genscript: A01736); Biotinylated Protein L (Genscript: M0097); Streptavidin-APC conjugate (Molecular probes: SA1005); APC-conjugated anti-MYC antibody (R&D systems: IC3696A); and Polyethylenimine (Polysciences Inc: 23966) were obtained from the indicated sources.

Techniques: Stable Transfection, Expressing, Incubation, Flow Cytometry, Staining

Journal: Scientific Reports

Article Title: A novel luciferase-based assay for the detection of Chimeric Antigen Receptors

doi: 10.1038/s41598-018-38258-z

Figure Lengend Snippet: ( A ) Non-transduced JNG cells and JNG cells stably expressing the indicated BCMA-CARs (Centyrin, J6MO, and BB-CAR-02) and a CD19-CAR (FMC63) were incubated with 100 µl of supernatant containing the BCMA-ECD-Nluc fusion protein for 45 minutes on ice. After incubation, cells were washed 5 times and assayed for luminescence. ( B ) The assay was performed as described in ( A ) except that CD19-ECD-Nluc fusion protein was used. ( C ) JNG cells stably expressing the indicated CARs were incubated with an APC-conjugated MYC antibody for 45 minutes on ice. After incubation, cells were washed twice and analyzed by flow cytometry. ( D ) JNG cells stably expressing the indicated CARs were stained with Protein L reagent as described in methods section followed by flow cytometry analysis. The values shown are mean ± SE of a representative of at least two experiments performed in duplicate.

Article Snippet: Strep-Tactin-APC (IBA-lifescience: Cat# 6-5010-001); Anti-Flag-FITC (Sigma: F4049); Anti-Strep-tagII-FITC (Genscript: A01736); Biotinylated Protein L (Genscript: M0097); Streptavidin-APC conjugate (Molecular probes: SA1005); APC-conjugated anti-MYC antibody (R&D systems: IC3696A); and Polyethylenimine (Polysciences Inc: 23966) were obtained from the indicated sources.

Techniques: Stable Transfection, Expressing, Incubation, Flow Cytometry, Staining

Journal: Scientific Reports

Article Title: A novel luciferase-based assay for the detection of Chimeric Antigen Receptors

doi: 10.1038/s41598-018-38258-z

Figure Lengend Snippet: ( A ) Human primary T cells transduced with a CD19-CAR and parental cells were stained with purified CD19-ECD fusion protein labelled with Alexa Fluor- 647 (AF647). Alternatively, the cells were labeled with Biotinylated Protein L followed by APC-conjugated Streptavidin. After washes, cells were analyzed by flow cytometry. ( B ) Human primary T cells expressing a CD19-CAR or a CD33-CAR were incubated with 100 µl of supernatants containing the epitope-tagged CD19- or CD33-ECD fusion proteins for 45 minutes on ice. After incubation, cells were washed 3 times and incubated with the indicated anti-tag antibodies for 45 minutes. After 2 washes, cells were analyzed by flow cytometry. A representative of two independent experiments is shown.

Article Snippet: Strep-Tactin-APC (IBA-lifescience: Cat# 6-5010-001); Anti-Flag-FITC (Sigma: F4049); Anti-Strep-tagII-FITC (Genscript: A01736); Biotinylated Protein L (Genscript: M0097); Streptavidin-APC conjugate (Molecular probes: SA1005); APC-conjugated anti-MYC antibody (R&D systems: IC3696A); and Polyethylenimine (Polysciences Inc: 23966) were obtained from the indicated sources.

Techniques: Transduction, Staining, Purification, Labeling, Flow Cytometry, Expressing, Incubation