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human cytokine duoset elisa kits  (R&D Systems)


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    R&D Systems human cytokine duoset elisa kits
    Isolation of PD-L1 variants with improved PD-1 binding affinity. ( A ) Flow cytometry analysis comparing PD-1 binding to yeast-displayed wild-type PD-L1 fused to Aga2 at either the N-terminus or C-terminus. The C-terminal fusion format resulted in significantly superior receptor engagement. ( B ) Crystal structure of the PD-1/PD-L1 complex (PDB ID: 3BIK), highlighting the N- and C-termini of PD-L1 relevant to display orientation. ( C ) FACS density plots showing progressive enrichment of PD-L1 variants with enhanced PD-1 binding over successive rounds of library screening. Yeast cells were co-stained with anti-FLAG-Alexa Fluor 647 and PD-1-GST-Alexa Fluor 488 probes. ( D ) <t>ELISA-based</t> quantification of PD-1 binding by seven affinity-matured PD-L1 variants (Y2–Y9), in comparison with wild-type PD-L1 and a previously reported variant (L3B3)
    Human Cytokine Duoset Elisa Kits, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 252 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+cytokine+duoset+elisa+kits/Human+Renin+DuoSet+ELISA/pmc12679730-61-11-19
    Average 95 stars, based on 252 article reviews
    human cytokine duoset elisa kits - by Bioz Stars, 2026-11
    95/100 stars

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    1) Product Images from "Directed evolution–driven reprogramming of PD-L1 for compact and tunable checkpoint modulation"

    Article Title: Directed evolution–driven reprogramming of PD-L1 for compact and tunable checkpoint modulation

    Journal: Journal of Biological Engineering

    doi: 10.1186/s13036-025-00577-x

    Isolation of PD-L1 variants with improved PD-1 binding affinity. ( A ) Flow cytometry analysis comparing PD-1 binding to yeast-displayed wild-type PD-L1 fused to Aga2 at either the N-terminus or C-terminus. The C-terminal fusion format resulted in significantly superior receptor engagement. ( B ) Crystal structure of the PD-1/PD-L1 complex (PDB ID: 3BIK), highlighting the N- and C-termini of PD-L1 relevant to display orientation. ( C ) FACS density plots showing progressive enrichment of PD-L1 variants with enhanced PD-1 binding over successive rounds of library screening. Yeast cells were co-stained with anti-FLAG-Alexa Fluor 647 and PD-1-GST-Alexa Fluor 488 probes. ( D ) ELISA-based quantification of PD-1 binding by seven affinity-matured PD-L1 variants (Y2–Y9), in comparison with wild-type PD-L1 and a previously reported variant (L3B3)
    Figure Legend Snippet: Isolation of PD-L1 variants with improved PD-1 binding affinity. ( A ) Flow cytometry analysis comparing PD-1 binding to yeast-displayed wild-type PD-L1 fused to Aga2 at either the N-terminus or C-terminus. The C-terminal fusion format resulted in significantly superior receptor engagement. ( B ) Crystal structure of the PD-1/PD-L1 complex (PDB ID: 3BIK), highlighting the N- and C-termini of PD-L1 relevant to display orientation. ( C ) FACS density plots showing progressive enrichment of PD-L1 variants with enhanced PD-1 binding over successive rounds of library screening. Yeast cells were co-stained with anti-FLAG-Alexa Fluor 647 and PD-1-GST-Alexa Fluor 488 probes. ( D ) ELISA-based quantification of PD-1 binding by seven affinity-matured PD-L1 variants (Y2–Y9), in comparison with wild-type PD-L1 and a previously reported variant (L3B3)

    Techniques Used: Isolation, Binding Assay, Flow Cytometry, Library Screening, Staining, Enzyme-linked Immunosorbent Assay, Comparison, Variant Assay

    Identification and binding characterization of engineered PD-L1 variants with enhanced affinity for PD-1. ( A ) Amino acid sequence alignment of affinity-matured PD-L1 variants in comparison to wild-type PD-L1. Two conserved mutations, N45D and V50M, are found recurrently among the top-performing variants. ( B ) ELISA analysis of PD-1 binding by selected PD-L1 variants (Y2, Y3, Y6), a single mutant (D), and a double mutant (DM), compared with wild-type PD-L1 and L3B3. ( C ) Comparative ELISA of PD-1 binding for the wild-type, L3B3, L3C7c, and DM variants, highlighting the superior affinity of the DM variant. ( D ) Kinetic parameters derived from bio-layer interferometry (BLI), including association ( k a ), dissociation ( k d ), and equilibrium dissociation constants ( K D ), confirming the DM variant as the highest-affinity binder. The BLI assay data, including fitted kinetic constants and their associated error values, are presented in Supplementary Figure
    Figure Legend Snippet: Identification and binding characterization of engineered PD-L1 variants with enhanced affinity for PD-1. ( A ) Amino acid sequence alignment of affinity-matured PD-L1 variants in comparison to wild-type PD-L1. Two conserved mutations, N45D and V50M, are found recurrently among the top-performing variants. ( B ) ELISA analysis of PD-1 binding by selected PD-L1 variants (Y2, Y3, Y6), a single mutant (D), and a double mutant (DM), compared with wild-type PD-L1 and L3B3. ( C ) Comparative ELISA of PD-1 binding for the wild-type, L3B3, L3C7c, and DM variants, highlighting the superior affinity of the DM variant. ( D ) Kinetic parameters derived from bio-layer interferometry (BLI), including association ( k a ), dissociation ( k d ), and equilibrium dissociation constants ( K D ), confirming the DM variant as the highest-affinity binder. The BLI assay data, including fitted kinetic constants and their associated error values, are presented in Supplementary Figure

    Techniques Used: Binding Assay, Sequencing, Comparison, Enzyme-linked Immunosorbent Assay, Mutagenesis, Variant Assay, Derivative Assay

    PD-1 binding inhibition and T cell activation induced by Fc-fused PD-L1 variants. ( A and B ) Competitive ELISA demonstrating the ability of Fc-fused PD-L1 variants to inhibit PD-1 binding to immobilized ligands. Increasing concentrations of Fc-fused wild-type and DM PD-L1 variants were tested for their ability to block PD-1 interaction with ( A ) plate-bound PD-L1-Fc or ( B ) PD-L2-Fc. Absorbance at 450 nm reflects residual PD-1 binding after competition, with error bars indicating variation between duplicate samples. ( C and D ) Mixed lymphocyte reaction (MLR) assay evaluating the immunostimulatory capacity of Fc-fused PD-L1 variants. CD4⁺ T cells were co-cultured with mature dendritic cells in the presence of the indicated Fc-fused ligands. Cytokine levels in the supernatants were quantified by ELISA. ( C ) Interferon-γ (IFN-γ) and (D) interleukin-2 (IL-2) production, expressed in pg/mL. Statistical significance for cytokine secretion was assessed using one-way ANOVA as follows: p > 0.05 (ns, not significant), p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***)
    Figure Legend Snippet: PD-1 binding inhibition and T cell activation induced by Fc-fused PD-L1 variants. ( A and B ) Competitive ELISA demonstrating the ability of Fc-fused PD-L1 variants to inhibit PD-1 binding to immobilized ligands. Increasing concentrations of Fc-fused wild-type and DM PD-L1 variants were tested for their ability to block PD-1 interaction with ( A ) plate-bound PD-L1-Fc or ( B ) PD-L2-Fc. Absorbance at 450 nm reflects residual PD-1 binding after competition, with error bars indicating variation between duplicate samples. ( C and D ) Mixed lymphocyte reaction (MLR) assay evaluating the immunostimulatory capacity of Fc-fused PD-L1 variants. CD4⁺ T cells were co-cultured with mature dendritic cells in the presence of the indicated Fc-fused ligands. Cytokine levels in the supernatants were quantified by ELISA. ( C ) Interferon-γ (IFN-γ) and (D) interleukin-2 (IL-2) production, expressed in pg/mL. Statistical significance for cytokine secretion was assessed using one-way ANOVA as follows: p > 0.05 (ns, not significant), p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***)

    Techniques Used: Binding Assay, Inhibition, Activation Assay, Competitive ELISA, Blocking Assay, Mlr Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Directed evolution–driven reprogramming of PD-L1 for compact and tunable checkpoint modulation
    Article Snippet: RPMI 1640 medium, fetal bovine serum (FBS), antibiotic-antimycotic, sodium pyruvate, MEM non-essential amino acids solution, FreeStyle 293 expression medium, and Alexa Fluor 488 protein labeling kit were supplied by Thermo Fisher Scientific (Waltham, MA, USA). .. Interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and human cytokine DuoSet ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA). .. Interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were obtained from Peprotech (Cranbury, NJ, USA).



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    Image Search Results


    Isolation of PD-L1 variants with improved PD-1 binding affinity. ( A ) Flow cytometry analysis comparing PD-1 binding to yeast-displayed wild-type PD-L1 fused to Aga2 at either the N-terminus or C-terminus. The C-terminal fusion format resulted in significantly superior receptor engagement. ( B ) Crystal structure of the PD-1/PD-L1 complex (PDB ID: 3BIK), highlighting the N- and C-termini of PD-L1 relevant to display orientation. ( C ) FACS density plots showing progressive enrichment of PD-L1 variants with enhanced PD-1 binding over successive rounds of library screening. Yeast cells were co-stained with anti-FLAG-Alexa Fluor 647 and PD-1-GST-Alexa Fluor 488 probes. ( D ) ELISA-based quantification of PD-1 binding by seven affinity-matured PD-L1 variants (Y2–Y9), in comparison with wild-type PD-L1 and a previously reported variant (L3B3)

    Journal: Journal of Biological Engineering

    Article Title: Directed evolution–driven reprogramming of PD-L1 for compact and tunable checkpoint modulation

    doi: 10.1186/s13036-025-00577-x

    Figure Lengend Snippet: Isolation of PD-L1 variants with improved PD-1 binding affinity. ( A ) Flow cytometry analysis comparing PD-1 binding to yeast-displayed wild-type PD-L1 fused to Aga2 at either the N-terminus or C-terminus. The C-terminal fusion format resulted in significantly superior receptor engagement. ( B ) Crystal structure of the PD-1/PD-L1 complex (PDB ID: 3BIK), highlighting the N- and C-termini of PD-L1 relevant to display orientation. ( C ) FACS density plots showing progressive enrichment of PD-L1 variants with enhanced PD-1 binding over successive rounds of library screening. Yeast cells were co-stained with anti-FLAG-Alexa Fluor 647 and PD-1-GST-Alexa Fluor 488 probes. ( D ) ELISA-based quantification of PD-1 binding by seven affinity-matured PD-L1 variants (Y2–Y9), in comparison with wild-type PD-L1 and a previously reported variant (L3B3)

    Article Snippet: Interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and human cytokine DuoSet ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA).

    Techniques: Isolation, Binding Assay, Flow Cytometry, Library Screening, Staining, Enzyme-linked Immunosorbent Assay, Comparison, Variant Assay

    Identification and binding characterization of engineered PD-L1 variants with enhanced affinity for PD-1. ( A ) Amino acid sequence alignment of affinity-matured PD-L1 variants in comparison to wild-type PD-L1. Two conserved mutations, N45D and V50M, are found recurrently among the top-performing variants. ( B ) ELISA analysis of PD-1 binding by selected PD-L1 variants (Y2, Y3, Y6), a single mutant (D), and a double mutant (DM), compared with wild-type PD-L1 and L3B3. ( C ) Comparative ELISA of PD-1 binding for the wild-type, L3B3, L3C7c, and DM variants, highlighting the superior affinity of the DM variant. ( D ) Kinetic parameters derived from bio-layer interferometry (BLI), including association ( k a ), dissociation ( k d ), and equilibrium dissociation constants ( K D ), confirming the DM variant as the highest-affinity binder. The BLI assay data, including fitted kinetic constants and their associated error values, are presented in Supplementary Figure

    Journal: Journal of Biological Engineering

    Article Title: Directed evolution–driven reprogramming of PD-L1 for compact and tunable checkpoint modulation

    doi: 10.1186/s13036-025-00577-x

    Figure Lengend Snippet: Identification and binding characterization of engineered PD-L1 variants with enhanced affinity for PD-1. ( A ) Amino acid sequence alignment of affinity-matured PD-L1 variants in comparison to wild-type PD-L1. Two conserved mutations, N45D and V50M, are found recurrently among the top-performing variants. ( B ) ELISA analysis of PD-1 binding by selected PD-L1 variants (Y2, Y3, Y6), a single mutant (D), and a double mutant (DM), compared with wild-type PD-L1 and L3B3. ( C ) Comparative ELISA of PD-1 binding for the wild-type, L3B3, L3C7c, and DM variants, highlighting the superior affinity of the DM variant. ( D ) Kinetic parameters derived from bio-layer interferometry (BLI), including association ( k a ), dissociation ( k d ), and equilibrium dissociation constants ( K D ), confirming the DM variant as the highest-affinity binder. The BLI assay data, including fitted kinetic constants and their associated error values, are presented in Supplementary Figure

    Article Snippet: Interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and human cytokine DuoSet ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA).

    Techniques: Binding Assay, Sequencing, Comparison, Enzyme-linked Immunosorbent Assay, Mutagenesis, Variant Assay, Derivative Assay

    PD-1 binding inhibition and T cell activation induced by Fc-fused PD-L1 variants. ( A and B ) Competitive ELISA demonstrating the ability of Fc-fused PD-L1 variants to inhibit PD-1 binding to immobilized ligands. Increasing concentrations of Fc-fused wild-type and DM PD-L1 variants were tested for their ability to block PD-1 interaction with ( A ) plate-bound PD-L1-Fc or ( B ) PD-L2-Fc. Absorbance at 450 nm reflects residual PD-1 binding after competition, with error bars indicating variation between duplicate samples. ( C and D ) Mixed lymphocyte reaction (MLR) assay evaluating the immunostimulatory capacity of Fc-fused PD-L1 variants. CD4⁺ T cells were co-cultured with mature dendritic cells in the presence of the indicated Fc-fused ligands. Cytokine levels in the supernatants were quantified by ELISA. ( C ) Interferon-γ (IFN-γ) and (D) interleukin-2 (IL-2) production, expressed in pg/mL. Statistical significance for cytokine secretion was assessed using one-way ANOVA as follows: p > 0.05 (ns, not significant), p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***)

    Journal: Journal of Biological Engineering

    Article Title: Directed evolution–driven reprogramming of PD-L1 for compact and tunable checkpoint modulation

    doi: 10.1186/s13036-025-00577-x

    Figure Lengend Snippet: PD-1 binding inhibition and T cell activation induced by Fc-fused PD-L1 variants. ( A and B ) Competitive ELISA demonstrating the ability of Fc-fused PD-L1 variants to inhibit PD-1 binding to immobilized ligands. Increasing concentrations of Fc-fused wild-type and DM PD-L1 variants were tested for their ability to block PD-1 interaction with ( A ) plate-bound PD-L1-Fc or ( B ) PD-L2-Fc. Absorbance at 450 nm reflects residual PD-1 binding after competition, with error bars indicating variation between duplicate samples. ( C and D ) Mixed lymphocyte reaction (MLR) assay evaluating the immunostimulatory capacity of Fc-fused PD-L1 variants. CD4⁺ T cells were co-cultured with mature dendritic cells in the presence of the indicated Fc-fused ligands. Cytokine levels in the supernatants were quantified by ELISA. ( C ) Interferon-γ (IFN-γ) and (D) interleukin-2 (IL-2) production, expressed in pg/mL. Statistical significance for cytokine secretion was assessed using one-way ANOVA as follows: p > 0.05 (ns, not significant), p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***)

    Article Snippet: Interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and human cytokine DuoSet ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA).

    Techniques: Binding Assay, Inhibition, Activation Assay, Competitive ELISA, Blocking Assay, Mlr Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

    CXCL-8 secretion varied with glycan alterations. (A) Experimental workflow for assessing pro-inflammatory cytokine secretion of glycosylation mutants compared to wildtype H. pylori . Image was created using BioRender. (B) The glycoprotein mutant with elaborated LPS structures, Δ579, induced increased CXCL-8 secretion from gastric epithelial cells relative to wildtype. (C) The LPS mutant, Δwzk, induced decreased CXCL-8 secretion from gastric epithelial cells relative to wildtype. (D–I) Variable impact on CXCL-8 secretion was observed with glycoprotein mutants Δ580 and Δ1179, and LPS mutant, ΔwaaL, compared to wildtype depending on the coculture experiment. Error bars reflect technical replicates. Tukey’s multiple comparison test one-way ANOVA was used. (**P< 0.01, ***P< 0.001, ****P< 0.0001, ns, not significant). Data are representative of independent replicate experiments (n > 3) that exhibited the same findings.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Helicobacter pylori glycan biosynthesis modulates host immune cell recognition and response

    doi: 10.3389/fcimb.2024.1377077

    Figure Lengend Snippet: CXCL-8 secretion varied with glycan alterations. (A) Experimental workflow for assessing pro-inflammatory cytokine secretion of glycosylation mutants compared to wildtype H. pylori . Image was created using BioRender. (B) The glycoprotein mutant with elaborated LPS structures, Δ579, induced increased CXCL-8 secretion from gastric epithelial cells relative to wildtype. (C) The LPS mutant, Δwzk, induced decreased CXCL-8 secretion from gastric epithelial cells relative to wildtype. (D–I) Variable impact on CXCL-8 secretion was observed with glycoprotein mutants Δ580 and Δ1179, and LPS mutant, ΔwaaL, compared to wildtype depending on the coculture experiment. Error bars reflect technical replicates. Tukey’s multiple comparison test one-way ANOVA was used. (**P< 0.01, ***P< 0.001, ****P< 0.0001, ns, not significant). Data are representative of independent replicate experiments (n > 3) that exhibited the same findings.

    Article Snippet: Human Cytokine DuoSet Enzyme Linked Immunoassay (ELISA) kits (R&D Systems) were utilized to detect relative concentrations of CXCL-8 (IL-8), IL-10, TNF- α IL-6, or IL-1 β conditioned media samples.

    Techniques: Glycoproteomics, Mutagenesis, Comparison

    Immature dendritic cell cytokine secretion changed when challenged by H. pylori glycosylation mutants. (A) Experimental workflow used to assess TNF-α, IL-6, IL-1β, and IL-10 secretion from iDCs following challenge for 24-hours with glycosylation mutants or wildtype bacteria. Image was created using BioRender. Datasets from two different iDC challenge experiments are shown, with (B–E) collected in one experiment and (F–I) collected in a second experiment. Δ580.1 and Δ 580.2, as well as Δ1179.1 and Δ1179.2, represent different freeze lots of the same strain. Error bars reflect technical replicates. Tukey’s multiple comparison test one-way ANOVA was used. (*P< 0.05, **P< 0.01, ***P< 0.001, ****P< 0.0001, ns, not significant).

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Helicobacter pylori glycan biosynthesis modulates host immune cell recognition and response

    doi: 10.3389/fcimb.2024.1377077

    Figure Lengend Snippet: Immature dendritic cell cytokine secretion changed when challenged by H. pylori glycosylation mutants. (A) Experimental workflow used to assess TNF-α, IL-6, IL-1β, and IL-10 secretion from iDCs following challenge for 24-hours with glycosylation mutants or wildtype bacteria. Image was created using BioRender. Datasets from two different iDC challenge experiments are shown, with (B–E) collected in one experiment and (F–I) collected in a second experiment. Δ580.1 and Δ 580.2, as well as Δ1179.1 and Δ1179.2, represent different freeze lots of the same strain. Error bars reflect technical replicates. Tukey’s multiple comparison test one-way ANOVA was used. (*P< 0.05, **P< 0.01, ***P< 0.001, ****P< 0.0001, ns, not significant).

    Article Snippet: Human Cytokine DuoSet Enzyme Linked Immunoassay (ELISA) kits (R&D Systems) were utilized to detect relative concentrations of CXCL-8 (IL-8), IL-10, TNF- α IL-6, or IL-1 β conditioned media samples.

    Techniques: Glycoproteomics, Bacteria, Comparison

    The glycocalyx was probed to assess how Lewis Y expression and LPS elaboration correlate to cytokine secretion by immune cells. (A) Western blot analysis reveals robust Lewis Y expression in Δ579 and Δ580.2. The strain samples used in these experiments correspond to those used in iDC challenge experiments for the dataset shown in <xref ref-type= Figure 5 . (B, C) Analysis of LPS by gel electrophoresis reveals that Δwzk and ΔwaaL synthesize only truncated, low molecular weight LPS relative to wildtype H. pylori , whereas Δ579 appears to synthesize LPS with somewhat more prominent bands at higher molecular weights (e.g., >29kDa) than wildtype H. pylori . (B) The strain samples used in these experiments correspond to those used in dendritic cell challenge experiments for the dataset shown in Figure 5 . Δ580.1 and Δ 580.2 represent different freeze lots of the same strain. (C) Analysis of LPS by gel electrophoresis for strain samples used in dendritic cell challenge experiments for the dataset shown in Supplementary Figure S3 . Δ1179.1 and Δ1179.2 represent different freeze lots of the same strain and appear to have comparable LPS fingerprints. " width="100%" height="100%">

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Helicobacter pylori glycan biosynthesis modulates host immune cell recognition and response

    doi: 10.3389/fcimb.2024.1377077

    Figure Lengend Snippet: The glycocalyx was probed to assess how Lewis Y expression and LPS elaboration correlate to cytokine secretion by immune cells. (A) Western blot analysis reveals robust Lewis Y expression in Δ579 and Δ580.2. The strain samples used in these experiments correspond to those used in iDC challenge experiments for the dataset shown in Figure 5 . (B, C) Analysis of LPS by gel electrophoresis reveals that Δwzk and ΔwaaL synthesize only truncated, low molecular weight LPS relative to wildtype H. pylori , whereas Δ579 appears to synthesize LPS with somewhat more prominent bands at higher molecular weights (e.g., >29kDa) than wildtype H. pylori . (B) The strain samples used in these experiments correspond to those used in dendritic cell challenge experiments for the dataset shown in Figure 5 . Δ580.1 and Δ 580.2 represent different freeze lots of the same strain. (C) Analysis of LPS by gel electrophoresis for strain samples used in dendritic cell challenge experiments for the dataset shown in Supplementary Figure S3 . Δ1179.1 and Δ1179.2 represent different freeze lots of the same strain and appear to have comparable LPS fingerprints.

    Article Snippet: Human Cytokine DuoSet Enzyme Linked Immunoassay (ELISA) kits (R&D Systems) were utilized to detect relative concentrations of CXCL-8 (IL-8), IL-10, TNF- α IL-6, or IL-1 β conditioned media samples.

    Techniques: Expressing, Western Blot, Nucleic Acid Electrophoresis, Molecular Weight