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Kir2dl3 fc chimera

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KIR2DL3-Fc chimera is a recombinant protein that consists of the extracellular domain of the human killer cell immunoglobulin-like receptor KIR2DL3 fused to the Fc portion of human IgG1. This protein can be used in various immunological assays and research applications.

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Lab products found in correlation

Kir2dl2 fc, by R&D Systems (2 mentions) Protein a alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Lsrfortessa, by BD (1 mentions)

Close spelling variants of this product

KIR2DL3 (2 citations)

4 protocols using kir2dl3 fc chimera

1

KIR2DL3-Fc Binding Assay for Peptide-Pulsed Cells

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For KIR-Fc staining 221-TAPko-HLA-C*03:04 cells pulsed with YFV/NSA2A4−13-, HIV/Gag296−303- or HCV/Core136−144-peptides were stained using KIR2DL3-Fc chimera (R&D) for 1 h on ice. After two washing steps with ice-cold 2% FBS/PBS cells were incubated with 1.25 μL of goat anti-hIgG(Fc)-PE antibody (Fisher Scientific). After washing with ice-cold 2% FBS/PBS cells were fixed with Fixation Buffer and analyzed using flow cytometry (BD LSR Fortessa). Measurement of KIR-Fc binding was assessed as percentage of PE-positive cells.
Ziegler M.C., Grañana F.B., Garcia-Beltran W.F., Schulze zur Wiesch J., Hoffmann C., Rechtien A., Lunemann S, & Altfeld M. (2018). Stable Frequencies of HLA-C*03:04/Peptide-Binding KIR2DL2/3+ Natural Killer Cells Following Vaccination. Frontiers in Immunology, 9, 2361.
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2

KIR-Fc Chimera Binding Assay

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KIR2D2L2‐IgG and KIR2DL3‐IgG fusion constructs (KIR2DL2‐Fc & KIR2DL3‐Fc Chimera; R&D Systems) were conjugated with protein A Alexa Fluor 488 (Invitrogen) at a molar ratio of 6:1. A total of 1 × 106 721.174 cells were incubated with 10 μM peptide as described above and then stained with KIR‐Fc. After fixation in 4% w/v paraformaldehyde cells were analyzed by flow cytometry.
Cassidy S., Mukherjee S., Myint T.M., Mbiribindi B., North H., Traherne J., Mulder A., Claas F.H., Purbhoo M.A., Das J, & Khakoo S.I. (2014). Peptide selectivity discriminates NK cells from KIR2DL2‐ and KIR2DL3‐positive individuals. European Journal of Immunology, 45(2), 492-500.
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3

HLA-C*03:04 Stabilization and KIR Binding Assays

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HLA stabilization and KIR binding assays were performed as previously described [16 (link)]. In short, cells were washed with FBS free RPMI 1640 (R0) to remove any remaining foreign peptides from culturing in R10. Afterwards 721.221-ICP47-C*03:04, or 221-C*03:04-TAP1-KO cells were pulsed with 100μM of the respective HCV peptide for 20h at 26°C. Previously described peptides that stabilized HLA-C*03:04 expression GAVDPLLAL (GAL) and GAVDPLLKL (GKL) [17 (link)], were used as positive controls, whereas culturing in the absence of peptides was used as negative control. After peptide-pulsing, cells were stained using an anti-HLA-ABC antibody (clone W6/32), fixed in 4% paraformaldehyde and analyzed using flow cytometry. KIR binding was analyzed after 20h of HLA stabilization, as described above, with the respective peptides of interest. Cells were stained using KIR2DL3-Fc chimera (R&D), for 1h on ice, secondary staining with anti-human FC antibody was performed for 30min on ice. Afterwards cells were fixed and analyzed using flow cytometry.
Lunemann S., Martrus G., Hölzemer A., Chapel A., Ziegler M., Körner C., Beltran W.G., Carrington M., Wedemeyer H, & Altfeld M. (2016). Sequence variations in HCV core-derived epitopes alter binding of KIR2DL3 to HLA-C*03:04 and modulate NK cell function. Journal of hepatology, 65(2), 252-258.
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4

KIR2DL2-Fc and KIR2DL3-Fc Binding Assay

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KIR2D2L2-IgG and KIR2DL3-IgG fusion constructs (KIR2DL2-Fc & KIR2DL3-Fc Chimera; R&D Systems) were conjugated with protein A Alexa Fluor 488 (Invitrogen) at a molar ratio of 6:1. 1 × 106 721.174 cells were incubated with 10 μM peptide as described above and then stained with KIR-Fc. After fixation in 4% (w/v) paraformaldehyde cells were analyzed by flow cytometry.
Cassidy S., Mukherjee S., Myint T.M., Mbiribindi B., North H., Traherne J., Mulder A., Claas F.H., Purbhoo M.A., Das J, & Khakoo S.I. (2014). Peptide selectivity discriminates NK cells from KIR2DL2‐ and KIR2DL3‐positive individuals. European Journal of Immunology, 45(2), 492-500.
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