recombinant human igg2  (Millipore)

Journal: NPJ Parkinson's Disease

Article Title: Novel co-culture model of T cells and midbrain organoids for investigating neurodegeneration in Parkinson’s disease

doi: 10.1038/s41531-025-00882-8

Figure Lengend Snippet: Antibodies used for flow cytometry analyses

Article Snippet: IL-17 FITC , recombinant human IgG2 , Miltenyi Biotec , REA1063 , 130-118-242.

Techniques: Flow Cytometry, Recombinant

Journal: NPJ Parkinson's Disease

Article Title: Novel co-culture model of T cells and midbrain organoids for investigating neurodegeneration in Parkinson’s disease

doi: 10.1038/s41531-025-00882-8

Figure Lengend Snippet: Antibodies used in ICC

Article Snippet: IL-17 FITC , recombinant human IgG2 , Miltenyi Biotec , REA1063 , 130-118-242.

Techniques: Recombinant

Journal: STAR Protocols

Article Title: FlowLITE: A protocol to characterize and quantify total antibody isotypes in human plasma using flow cytometry

doi: 10.1016/j.xpro.2025.104200

Figure Lengend Snippet: Schematic of 8-bead FlowLITE assay (A) FlowLITE consists of fluorescent yellow beads coated with anti-human isotype antibodies (monoclonal mouse IgG1). These antibodies are specific to the isotype variations in the heavy chain constant region. The bound antibodies from the sample are revealed by a fluorescently conjugated antibody specific to the light chain of the captured human antibody. (B) FlowLITE can be used to distinguish human antibody isotypes. Isotypes are distinguished by color and structure in the schematic.

Article Snippet: Purified Recombinant Human IgG2 Kappa (clone AbD18705_hIgG2); starting concentration: 0.01 mg/mL , Bio-Rad , Cat#HCA193.

Techniques:

Journal: STAR Protocols

Article Title: FlowLITE: A protocol to characterize and quantify total antibody isotypes in human plasma using flow cytometry

doi: 10.1016/j.xpro.2025.104200

Figure Lengend Snippet: Analysis of human plasma using 3-bead FlowLITE assay (IgM, pan-IgG, and pan-IgA) (A) Gating strategy for discrimination of anti-type bead species based on intensity of yellow fluorescence, detected in the V5 channel on the Cytek Aurora. (B) Titration of purified human IgM, IgG, and IgA. A 2-fold dilution series of the purified antibody was performed, starting from a concentration of 0.01 mg/mL. Background subtraction was performed using a control bead incubated with flowLITE media. (C) Linear range of detection (9.8 to 5,000 ng/mL) and ordinary least squares regression output (performed in Excel). Antibody concentration is determined using the linear regression calculated for each anti-isotype bead. The regression is adjusted based on the starting concentration of antibody in the standard curve (0.01 mg/mL). This value is represented by a constant, C =– log 2 (0.01 mg / ml )–1. (D) 2-fold dilution series of healthy human plasma starting from a concentration of 1:250. (E) Inter-assay variability (across days and runs) of quantified concentrations of antibody in mg/dL of the same donor at an optimal dilution of 1:64000 (dilution step 9 as indicated by the red box in D) was assessed in triplicate. The concentrations of IgM, IgG, and IgA were calculated and presented as the geometric mean and standard error for the three trials.

Article Snippet: Purified Recombinant Human IgG2 Kappa (clone AbD18705_hIgG2); starting concentration: 0.01 mg/mL , Bio-Rad , Cat#HCA193.

Techniques: Clinical Proteomics, Fluorescence, Titration, Purification, Concentration Assay, Control, Incubation, Inter Assay

Journal: STAR Protocols

Article Title: FlowLITE: A protocol to characterize and quantify total antibody isotypes in human plasma using flow cytometry

doi: 10.1016/j.xpro.2025.104200

Figure Lengend Snippet: Optimal bead saturation determined by titration of anti-human isotype capture antibodies (A) Schematic diagram of bead loading as the concentration of anti-isotype monoclonal mouse IgG1 antibody monomers decreases. The concatenated plot illustrates the saturation of the bead as the capture antibody concentration decreases. The dashed line indicates the base fluorescence of the bead in the YG1 channel when the coated bead is incubated in FlowLITE media. (B) Anti-IgE bead load titration was performed using yellow peak 9 (Y9) from the 12-set yellow-labeled beads (Spherotech). MFI values plotted against anti-IgE antibody concentration starting from a molarity of 1.6 μM (40 pmol/25 μL). (C) Bead load titrations of bead species Y3 (IgG1), Y4 (IgG2), Y5 (IgG3), Y6(IgG4) conjugated to the denoted anti-IgG subtype antibodies.

Article Snippet: Purified Recombinant Human IgG2 Kappa (clone AbD18705_hIgG2); starting concentration: 0.01 mg/mL , Bio-Rad , Cat#HCA193.

Techniques: Titration, Concentration Assay, Fluorescence, Incubation, Labeling

Journal: STAR Protocols

Article Title: FlowLITE: A protocol to characterize and quantify total antibody isotypes in human plasma using flow cytometry

doi: 10.1016/j.xpro.2025.104200

Figure Lengend Snippet: Application of FlowLITE in 8-bead assay and quantification of IgG subclasses (A) Readout of an 8-bead FlowLITE assay for detection of isotype subclasses. (B) Pie chart representation of the MFI ratios of the antibody isotypes and subclasses in the human plasma of a healthy donor. (C) Linear regressions for the calibration curves of the different IgG subclasses utilizing purified kappa IgG antibodies (Bio-Rad). (D) IgG subclass calibration curve regression output and quantification of IgG subclasses in human plasma.

Article Snippet: Purified Recombinant Human IgG2 Kappa (clone AbD18705_hIgG2); starting concentration: 0.01 mg/mL , Bio-Rad , Cat#HCA193.

Techniques: Clinical Proteomics, Purification

Journal: STAR Protocols

Article Title: FlowLITE: A protocol to characterize and quantify total antibody isotypes in human plasma using flow cytometry

doi: 10.1016/j.xpro.2025.104200

Figure Lengend Snippet: Reporting the absolute count of antibody molecules with QuantiBrite (A) PE QuantiBrite beads were used for calibration. A linear regression was performed to correlate fluorescence with the number of PE molecules. (B) The QuantiBrite regression was used to calculate the concentration of IgG molecules in a purified antibody sample. A 2-fold dilution of purified human IgG (starting at 0.1 mg/mL) was assessed using an antigen-coated bead (FlowBEAT format) and a pan-IgG reveal. Fluorescence was converted to PE copies using the formula derived in (A), assuming a conjugation of 1 PE per reveal antibody.

Article Snippet: Purified Recombinant Human IgG2 Kappa (clone AbD18705_hIgG2); starting concentration: 0.01 mg/mL , Bio-Rad , Cat#HCA193.

Techniques: Fluorescence, Concentration Assay, Purification, Derivative Assay, Conjugation Assay

Journal: Scientific Reports

Article Title: Class switch towards spike protein-specific IgG4 antibodies after SARS-CoV-2 mRNA vaccination depends on prior infection history

doi: 10.1038/s41598-023-40103-x

Figure Lengend Snippet: The total spike-specific IgG and the total serum IgG levels in vaccinated and COVID-19 cohorts. Total spike-specific IgG levels were quantified by in-house ELISA, normalized to positive and negative serum samples ( a ). Total serum IgG levels were measured by nephelometry ( b ). Black and blue circles indicate mRNA and vector-vaccinated volunteers in the vaccinated cohort, while red circles represent the COVID-19 cohort. The horizontal solid lines indicate group medians, and the horizontal dashed lines indicate 25–75% percentiles. The dotted lines represent the reference ranges of total serum IgG by Siemens Healthcare Diagnostics Inc. The Kruskal–Wallis test followed by Dunn’s multiple comparisons test is presented for the vaccinated cohort. p < 0.05 was considered statistically significant.

Article Snippet: Native human IgG2 protein , abcam , ab90284.

Techniques: Enzyme-linked Immunosorbent Assay, Plasmid Preparation

Journal: Scientific Reports

Article Title: Class switch towards spike protein-specific IgG4 antibodies after SARS-CoV-2 mRNA vaccination depends on prior infection history

doi: 10.1038/s41598-023-40103-x

Figure Lengend Snippet: The spike-specific, the total serum IgG subclasses and the proportions of each spike-specific IgG subclass to their total serum IgG antibody subclass in the vaccinated ( a ) and the COVID-19 ( b ) cohorts. Levels of spike-specific IgG subclasses were quantified by in-house ELISA calibrated with purified human IgG 1, 2, 3 and 4 antibodies. Total serum IgG subclass concentrations were measured by nephelometry. Black and blue circles indicate mRNA and vector-vaccinated volunteers in the vaccinated cohort ( a ), while red circles represent the COVID-19 cohort ( b ). The horizontal solid lines indicate group medians, and the horizontal dashed lines indicate 25–75% percentiles. The dotted lines represent the reference ranges of total serum IgG subclass levels by Siemens Healthcare Diagnostics Inc. The values of the Kruskal–Wallis test followed by Dunn’s multiple comparisons test are presented in the vaccinated cohort. p < 0.05 was considered statistically significant.

Article Snippet: Native human IgG2 protein , abcam , ab90284.

Techniques: Enzyme-linked Immunosorbent Assay, Purification, Plasmid Preparation

Journal: Scientific Reports

Article Title: Class switch towards spike protein-specific IgG4 antibodies after SARS-CoV-2 mRNA vaccination depends on prior infection history

doi: 10.1038/s41598-023-40103-x

Figure Lengend Snippet: Heatmaps representing the log mean fold change values of spike-specific and total serum IgG subclasses in three follow-up mRNA vaccinated groups between two sampling time points after booster vaccination. The fold changes of spike-specific and total serum IgG subclasses and the proportions of each spike-specific to total serum IgG levels were visualized in heatmaps by the logarithmic values of mean ratios between the second and the first sampling time points. Decreasing trends are shown between 0 and (− 1) and increasing trends were plotted between 0 and 1. The constant level (no change) is indicated as 0.

Article Snippet: Native human IgG2 protein , abcam , ab90284.

Techniques: Sampling

Journal: Scientific Reports

Article Title: Class switch towards spike protein-specific IgG4 antibodies after SARS-CoV-2 mRNA vaccination depends on prior infection history

doi: 10.1038/s41598-023-40103-x

Figure Lengend Snippet: Contribution of each spike-specific IgG subclass to the sum of all spike-specific IgG antibody levels. The percentages of each spike-specific IgG subclass relative to the sum of spike-specific IgG levels are shown for the vaccinated and COVID-19 cohorts. The numbers represent the mean percentages of individuals calculated by dividing each spike specific IgG levels with the sum of all spike specific IgG levels. Schematic representation of spike-specific IgG1%, IgG2%, IgG3% and IgG4% are shown in blue, orange, grey and yellow sectors, respectively. The size of the pie charts indicates the levels of all spike-specific IgG antibodies from Fig. .

Article Snippet: Native human IgG2 protein , abcam , ab90284.

Techniques:

Journal: Scientific Reports

Article Title: Class switch towards spike protein-specific IgG4 antibodies after SARS-CoV-2 mRNA vaccination depends on prior infection history

doi: 10.1038/s41598-023-40103-x

Figure Lengend Snippet: Key resources.

Article Snippet: Native human IgG2 protein , abcam , ab90284.

Techniques: Recombinant, Software

Journal: iScience

Article Title: Both COVID-19 infection and vaccination induce high-affinity cross-clade responses to SARS-CoV-2 variants

doi: 10.1016/j.isci.2022.104766

Figure Lengend Snippet: Study design and experimental approach (A) Flowchart for inclusion and exclusion into the study. 41 samples were included in the analysis, representing different patient groups. (B) Violin boxplot showing the distribution of IgG p(EC 50 ) values against the SARS-CoV-2 spike protein. A cutoff value of p(EC 50 ) ≥ 2 was chosen to define reactive samples. Blue dots represent samples of infected and/or vaccinated individuals. Yellow dots are non-infected and non-vaccinated negative controls.

Article Snippet: Recombinant human IgG2 (for ELISA), Human myeloma, 2.5 μg/mL , EMD Millipore , AG504; RRID: AB_97839.

Techniques: Infection

Journal: iScience

Article Title: Both COVID-19 infection and vaccination induce high-affinity cross-clade responses to SARS-CoV-2 variants

doi: 10.1016/j.isci.2022.104766

Figure Lengend Snippet: Characterization of affinity of SARS-CoV-2 antibodies to WT, delta, and omicron RBD variants (A) 2D scatter plot with integrated density contours. All quantifiable data points reflecting K A (in M −1 ) and IgG concentration values (in M) are plotted. 95% confidence intervals for each point are colored in light red. Triangles denote patients receiving the REGN-COV cocktail. RBD variants: WT (grey), delta (blue), omicron (yellow). Dotted lines represent the measurements of the same patient sample against different RBD variants. (B) Bar graph displaying the percentages of quantifiable samples for WT (grey), delta (blue), and omicron (yellow) RBD variants. Comparisons were performed including all samples, samples excluding those treated with REGN-COV, and only those treated with REGN-COV. Fisher’s exact test displayed no significant differences, at α = 0.01. (C and D) Boxplot analysis of K A values (C) and IgG concentrations (D) for WT, delta, and omicron RBD variants. (E) To employ a combined score of binding affinity ( K A ) and IgG concentration, we utilized the product K A x IgG concentration. (C-E): Colors denote treatment with REGN-COV (red) or absence of treatment (blue). Kruskal-Wallis (KW) with post-hoc Wilcoxon rank sum test (WC) after Holm correction for multiple comparisons was used, with α = 0.01. None of the group-wise comparisons reached statistical significance.

Article Snippet: Recombinant human IgG2 (for ELISA), Human myeloma, 2.5 μg/mL , EMD Millipore , AG504; RRID: AB_97839.

Techniques: Concentration Assay, Binding Assay

Journal: iScience

Article Title: Both COVID-19 infection and vaccination induce high-affinity cross-clade responses to SARS-CoV-2 variants

doi: 10.1016/j.isci.2022.104766

Figure Lengend Snippet: Correlation of affinity and IgG concentrations with clinically relevant parameters does not reveal clear differences between vaccinated and infected subgroups (A) 2D scatter plot with integrated density contours. All quantifiable data points reflecting K A (in M −1 ) and IgG concentration (in M) are plotted. 95% confidence intervals for each point are colored in light red. No distinct clusters were observed among patient groups infected/vaccinated (grey), infected/non-vaccinated (blue), non-infected/vaccinated (yellow); however, the REGN-COV-treated patients (red) clustered separately. (B) The same groups as in (A) depicted in a boxplot. Statistical analysis is shown in the graph. The RBD variants are color-coded. (C and D) No correlation between age (C) or sex (D) and K A x IgG concentration. (E) Although Kruskal-Wallis statistical testing indicates that the distributions are significantly different for different disease severities, pair-wise testing with the Wilcoxon rank sum test does not result in significance. (F) Trend towards increased K A x IgG concentration products in triple vaccinated individuals, without being statistically significant. (G) Same as (F) but additionally stratified according to vaccination/non-vaccination. A: Dotted lines represent the measurements of the same patient sample against different RBD variants. B, D-G: Kruskal-Wallis (KW) with post-hoc Wilcoxon rank sum test (WC) after Holm correction for multiple comparisons was used, with α = 0.01. C: The Pearson correlation coefficient was calculated. C-G: The patient groups are color-coded as in (A); however, the REGN-COV-treated patients were excluded from analyses.

Article Snippet: Recombinant human IgG2 (for ELISA), Human myeloma, 2.5 μg/mL , EMD Millipore , AG504; RRID: AB_97839.

Techniques: Infection, Concentration Assay

Journal: iScience

Article Title: Both COVID-19 infection and vaccination induce high-affinity cross-clade responses to SARS-CoV-2 variants

doi: 10.1016/j.isci.2022.104766

Figure Lengend Snippet: Analysis of antibody subtypes, correlation with MAAP parameters, and global feature profiling (A) Multiple heatmaps. Purple heatmap displaying p(EC 50 ) values (gradient) obtained with TRABI ELISA, for IgG, IgA, IgM, IgG1, IgG2, IgG3, and IgG4 antibodies. The SARS-CoV-2 WT spike ectodomain, the WT S1 domain, the WT S2 domain, the WT RBD, the delta RBD, and the omicron RBD variants as well as the nucleocapsid (NC) proteins were used. Orange heatmap displaying K A values, green heatmap displaying IgG concentration, grey heatmap displaying K A x IgG concentration obtained with MAAP against WT, delta, and omicron RBD variants. Additional heatmaps depict the age (red to blue), sex (orange: male; blue: female), number of vaccinations (yellow = 0, orange = 1, purple = 2, red = 3), treatment with the REGN-COV cocktail (green = TRUE), the strength of immunosuppression (none = light blue, light = turquoise, heavy = dark blue), the days post onset of infection (DPO) for patients with infection (pink), and disease severity (orange gradient). (B) Correlation between IgG p(EC 50 ) values of the spike ectodomain with K A , IgG concentrations, or the product K A x IgG concentration. (C) Principal component analysis using all TRABI ELISA values as input. The three plots represent different color-based clustering approaches.

Article Snippet: Recombinant human IgG2 (for ELISA), Human myeloma, 2.5 μg/mL , EMD Millipore , AG504; RRID: AB_97839.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Infection

Journal: iScience

Article Title: Both COVID-19 infection and vaccination induce high-affinity cross-clade responses to SARS-CoV-2 variants

doi: 10.1016/j.isci.2022.104766

Figure Lengend Snippet:

Article Snippet: Recombinant human IgG2 (for ELISA), Human myeloma, 2.5 μg/mL , EMD Millipore , AG504; RRID: AB_97839.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Purification