recombinant human rhil 15  (R&D Systems)

Journal: STAR Protocols

Article Title: Protocol for genomic editing in human resting primary NK cells and NK-92 cells via CRISPR-Cas9 ribonucleoproteins

doi: 10.1016/j.xpro.2024.103123

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Article Snippet: Human recombinant interleukin 15 protein (rhIL-15) , ImmunoTools , Cat# 11340155.

Techniques: Recombinant, DNA Extraction, Gene Knockout, Cell Isolation, Staining, Software, CRISPR, Flow Cytometry

Journal: PLoS ONE

Article Title: Interleukin-15 Plays a Central Role in Human Kidney Physiology and Cancer through the γc Signaling Pathway

doi: 10.1371/journal.pone.0031624

Figure Lengend Snippet: A) Scatchard's plot analysis: Effects of anti-IL-15Rβ and γc mAbs on IL-15 binding to RCC. For the IL-15 binding experiments, RCC7 cells were incubated with increasing concentrations of radioiodinated rIL-15 in presence or not of the following neutralizing mAbs: anti-IL-2Rβ and IL-2Rγ. The nonspecific cell binding was determined in the presence of radioiodinated rhIL-15 and a 100-fold excess of unlabeled rhIL-15. Cell-bound (B) and unbound (free, F) fractions were measured, and the specific bound fraction was calculated by subtracting the nonspecific binding from the cell-bound fraction. On the ordinate is plotted the ratio of the specific bound fraction (expressed in sites per cell) over the total concentration (bound plus free) of radioiodinated rIL-15 (expressed in pM). On the abscissa bound fraction (expressed in sites per cell). The high affinity specific IL-15 binding (Kd = 375 pM, 413 IL-15 binding sites per cell), which was completely abrogated by neutralizing antibody against the IL-2Rβ (inset) but not the γc chain, suggested the presence on RCC of an IL-15Rα/IL-2Rβ complex. B ) Detection of IL-15Rαβ complex by immunoprecipitation (IP) with anti-IL-15Rα (M161) or mouse IgG protein G-Sepharose-conjugate on total lysate (TL) of RCC7. Immunoprecipitated complexes were blotted either with anti-IL-2Rβ (sc-1046) and anti-IL-15Rα (sc-9172). C ) Stimulation for 10 and 40 min with physiologic (10 pg/mL) and supra-physiologic (10 ng/mL) concentrations of rhIL-15 induces the phosphorylation of MAPK ERK1/2 and IκBα in RPTEC and RCC7, whereas STAT5 activation was only observed in RPTEC. Histograms represent densitometry comparison of each factor normalized to β-actin in 3 different RCC (RCC5, RCC7, RCC8) and 3 RPTEC batches. * P<0.05 versus control, Mann-Whitney test. One experiment representative of a total of three is shown.

Article Snippet: Recombinant human non-glycosylated IL-15 (rhIL-15) and neutralizing anti-IL-2Rβ Mikβ1 mAb were obtained from ImmunoTools (Friesoythe, Germany) and Horseradish peroxidase (HRP)-conjugated and fluorescent-conjugated secondary Abs were from Jackson ImmunoResearch.

Techniques: Binding Assay, Incubation, Concentration Assay, Immunoprecipitation, Activation Assay, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Interleukin-15 Plays a Central Role in Human Kidney Physiology and Cancer through the γc Signaling Pathway

doi: 10.1371/journal.pone.0031624

Figure Lengend Snippet: A ) Immunofluorescence of cell–cell adhesion molecules show that IL-15 favors epithelial-mesenchymal transition (EMT) on RCC7, whereas it preserves the EMT commitment of RPTEC. The medium culture of RPTEC was not changed in order to induce the EMT process. Cells stimulated or not with 10 pg/ml of rhIL-15 for 5 days, were fixed and stained using standard immunofluorescence procedures with Abs against epithelial (cytokeratins and ZO-1) and mesenchymal markers (F-actin, ASO2 and vimentin). Similar results were obtained using different RCC (RCC5, RCC8) and RPTEC batches. B ) After 48 h, transfected RCC were treated for an additional 48 h with 10 pg/mL of rhIL-15 before evaluating the epithelial (cytokeratins) and mesenchymal (vimentin) markers expression by flow cytometry. RhIL-15 induced EMT was counterbalanced only in IL-2Rγ/JAK3 co-transfected RCC. Mean fluorescence intensity values for each marker are shown in each histogram. Results are representative of three experiments.

Article Snippet: Recombinant human non-glycosylated IL-15 (rhIL-15) and neutralizing anti-IL-2Rβ Mikβ1 mAb were obtained from ImmunoTools (Friesoythe, Germany) and Horseradish peroxidase (HRP)-conjugated and fluorescent-conjugated secondary Abs were from Jackson ImmunoResearch.

Techniques: Immunofluorescence, Staining, Transfection, Expressing, Flow Cytometry, Fluorescence, Marker