catalog numbers 9056 09 9070 09 9210 09 9200 09  (SouthernBiotech)

Journal: eBioMedicine

Article Title: Cross-binding antibodies capable of neutralising diverse hantaviruses are produced in response to Puumala virus infection

doi: 10.1016/j.ebiom.2025.106091

Figure Lengend Snippet: Schematic timeline of the symptoms associated with HFRS caused by PUUV infection. Following an incubation period of one to six weeks, HFRS symptoms manifest as five distinct phases: febrile, hypotensive, oliguric, polyuric, and convalescent. The average time for which each stage lasts is shown, in addition to the symptoms associated with each phase. Serum creatinine, a measure of kidney injury, is displayed, as is urine output as a marker of kidney function. IgM and IgG titres are also shown. The four time points at which patient sera were sampled are also shown in relation to each disease phase. Time point one (T1) was sampled six days post-symptom onset (PSO, range: 2–11 days PSO). Time point two (T2) was sampled 12 days PSO (range: 8–20 days PSO). Time point three (T3) was sampled 28 PSO (range: 21–64 days PSO). Time point four (T4) was sampled 197 days PSO (range: 180–256 days PSO). Adapted from Avsic-Zupanc et al. Created in BioRender. Clark, J. (2025) https://BioRender.com/p36u208 .

Article Snippet: When assaying other immunoglobulin subtypes, anti-human IgA HRP (Sigma–Aldrich, Cat# A0295, RRID: AB_257876 , 1:3000), anti-human IgM HRP (Southern Biotech, Cat# 2020-05, RRID: AB_2795603 , 1:3000), anti-human IgG1 (Southern Biotech, Cat# 9054-05, RRID: AB_2796627 , 1:5000), anti-human IgG2 (Southern Biotech, Cat# 9060-05, RRID: AB_2796633 , 1:5000), anti-human IgG3 (Southern Biotech, Cat# 9210-09, RRID: AB_2796701 , 1:5000), or anti-human IgG4 (Southern Biotech, Cat# 9200-05, RRID: AB_2796691 , 1:5000) were used.

Techniques: Infection, Incubation, Marker

Journal: eBioMedicine

Article Title: Cross-binding antibodies capable of neutralising diverse hantaviruses are produced in response to Puumala virus infection

doi: 10.1016/j.ebiom.2025.106091

Figure Lengend Snippet: Patient sera exhibit robust anti-PUUV IgG at all time points and cross-binding IgG by T4. ELISAs were carried out using patient sera and plates coated with ultracentrifuge-purified rVSV expressing the GnGc of A) PUUV, B) SEOV, C) DOBV, D) HTNV, E) SNV, or F) ANDV. Wild-type VSV G) was used as a negative control. IgG titres are shown as area under the curve (AUC) with the geometric mean and geometric standard deviation denoted by error bars. 11 age and sex matched healthy control sera were included as negative controls and an in-house, anti-PUUV monoclonal antibody was used as a positive control. AUC values were log10 transformed and statistical significance was interrogated via ordinary one-way ANOVA using Tukey's multiple comparisons test. Only comparisons with p ≤ 0.05 are shown. Limits of detection (LOD) were calculated for each antigen using the average + 3× standard deviations of the AUC of the negative control sera and are shown as dotted lines on each graph. The percentage of serum samples at each sampling time point with AUC values above the LOD are shown as pie charts, with the percentage of positive samples displayed in green. The n for each time point is displayed below each pie chart. H) Maximum likelihood phylogenetic tree of orthohantavirus M segment amino acid sequences. Clades are colour-coded according to the reservoir host species, with blue denoting viruses found in Arvicolinae, red viruses found in Sigmodontinae, and green viruses found in Murinae. Viruses included are: Puumala virus (PUUV), Prospect Hill virus (PHV), Tula virus (TULV), Andes virus (ANDV), Chocclo virus (CHOV), Black Creek Canal virus (BCCV), Sin Nombre virus (SNV), Dobrava-Belgrade virus (DOBV), Seoul virus (SEOV), and Hantaan virus (HTNV). Viruses that are included in this study are shown in bold. The scale bar represents amino acid substitutions per site, and bootstrap consensus support values are denoted at branches if less than 90.

Article Snippet: When assaying other immunoglobulin subtypes, anti-human IgA HRP (Sigma–Aldrich, Cat# A0295, RRID: AB_257876 , 1:3000), anti-human IgM HRP (Southern Biotech, Cat# 2020-05, RRID: AB_2795603 , 1:3000), anti-human IgG1 (Southern Biotech, Cat# 9054-05, RRID: AB_2796627 , 1:5000), anti-human IgG2 (Southern Biotech, Cat# 9060-05, RRID: AB_2796633 , 1:5000), anti-human IgG3 (Southern Biotech, Cat# 9210-09, RRID: AB_2796701 , 1:5000), or anti-human IgG4 (Southern Biotech, Cat# 9200-05, RRID: AB_2796691 , 1:5000) were used.

Techniques: Binding Assay, Purification, Expressing, Negative Control, Standard Deviation, Control, Positive Control, Transformation Assay, Sampling, Virus

Journal: eBioMedicine

Article Title: Cross-binding antibodies capable of neutralising diverse hantaviruses are produced in response to Puumala virus infection

doi: 10.1016/j.ebiom.2025.106091

Figure Lengend Snippet: The quantity of immunoglobulin subtypes changes between the acute and convalescent phases and is associated with a decrease in Fc effector function. ELISAs were carried out using patient sera to investigate A) anti-rVSV-PUUV IgM, B) anti-rVSV-PUUV IgA, C) anti-rVSV-SEOV IgM, and D) anti-rVSV-SEOV IgA. IgG subtype ELISAs were performed to investigate the quantities of anti-rVSV-PUUV E) IgG1, F) IgG2, G) IgG3, or H) IgG4 and anti-PUUV NP I) IgG1, J) IgG2, K) IgG3, or L) IgG4. M) A schematic of the luciferase-based reporter assay, which was utilised to characterise the effector activity promoted by patient sera. rVSV-PUUV-infected Huh7.5 cells were incubated with patient sera and reporter cells that express the FcγRIIIa receptor, coupled to a gene expression pathway that results in the production of luciferase. N) The FcγRIIIa activity of the sera are shown as AUCs, with the mean and standard deviation denoted by error bars. 11 age and sex matched healthy control sera were included as negative controls, and an in-house, anti-PUUV monoclonal antibody was used as a positive control. AUC values were log10 transformed and statistical significance was interrogated via ordinary one-way ANOVA using a Tukey's multiple comparisons test. Only comparisons with p ≤ 0.05 are shown. Limits of detection (LOD) are shown as dotted lines on each graph. The percentage of serum samples at each sampling time point with AUC values above the LOD are shown as pie charts with the percentage of positive samples displayed in green. The n for each time point is displayed below each pie chart.

Article Snippet: When assaying other immunoglobulin subtypes, anti-human IgA HRP (Sigma–Aldrich, Cat# A0295, RRID: AB_257876 , 1:3000), anti-human IgM HRP (Southern Biotech, Cat# 2020-05, RRID: AB_2795603 , 1:3000), anti-human IgG1 (Southern Biotech, Cat# 9054-05, RRID: AB_2796627 , 1:5000), anti-human IgG2 (Southern Biotech, Cat# 9060-05, RRID: AB_2796633 , 1:5000), anti-human IgG3 (Southern Biotech, Cat# 9210-09, RRID: AB_2796701 , 1:5000), or anti-human IgG4 (Southern Biotech, Cat# 9200-05, RRID: AB_2796691 , 1:5000) were used.

Techniques: Luciferase, Reporter Assay, Activity Assay, Infection, Incubation, Gene Expression, Standard Deviation, Control, Positive Control, Transformation Assay, Sampling

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: IgG reactivity to (A) purified protein derivative (PPD) from Mtb , (B) Mtb cytosolic protein, (C) Mtb culture filtrate, (D) Ag85A and Ag85B, (E) Mtb cell wall, (F) ESAT-6 and CFP-10, and (G) control respiratory syncytial virus (RSV) was determined by customized multiplex Luminex for each individual patient sample with TB over three serial dilutions. Each dot represents relative reactivity for one individual patient sample determined by the area under the curve (AUC), summarizing the median fluorescence intensity (MFI) from serial dilutions. Bars represent median and 95% confidence intervals for latent ( n = 18) and active ( n = 19) TB groups. The p values determined by a Mann-Whitney U test are marked by ^ for significance after adjustment for multiple comparisons by Benjamini-Hochberg.

Article Snippet: Mouse Anti-Human IgG2 Fc-PE HP6002 , SouthernBiotech , Cat#9070-09; RRID:AB_2796639.

Techniques: Purification, Control, Virus, Multiplex Assay, Luminex, Fluorescence, MANN-WHITNEY

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: (A) Representative chromatograms show patterns of individual glycoforms isolated from the Fc domain of antigen-specific and total bulk IgG in an individual patient with TB, determined by capillary electrophoresis. (B) Violin plots show the relative abundance of sialic acid, galactose, fucose, and bisecting N-acetylglucosamine (GlcNAc) across all individual glycoforms isolated from ESAT-6 and CFP-10 (red) and Mtb cell wall (blue) polyclonal IgG. Each dot represents an individual sample with latent ( n = 18) or active ( n = 19) TB. The median and interquartiles are shown. The dashed lines show the median RSV (green) and total bulk (purple) glycans. The p values determined by a Wilcoxon matched-pairs signed rank test are marked by ^ for significance after adjustment for multiple comparisons by Benjamini-Hochberg.

Article Snippet: Mouse Anti-Human IgG2 Fc-PE HP6002 , SouthernBiotech , Cat#9070-09; RRID:AB_2796639.

Techniques: Isolation, Electrophoresis

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: (A) Luminescence from the virulent Mtb H37Rv luminescent reporter strain relates to colony-forming units (CFUs). The significance was evaluated by Pearson correlation. (B) To test the effect of antibodies on intracellular Mtb , primary human monocyte-derived macrophages were first infected with the virulent Mtb H37Rv luminescent reporter strain, and the extracellular bacteria were washed away. Then, Mtb -infected primary human monocyte-derived macrophages (MOI = 1) were treated with IgG. Finally, the bacterial burden was quantified with >99% of detectable Mtb in the intracellular as compared to the extracellular medium supernatant compartment. The error bars represent the mean ± SEM. Significance was determined by a Wilcoxon matched-pairs signed rank test. (C) Daily Mtb luminescence measurements representing the median of endemic control, active TB, and latent TB samples are shown for one representative healthy donor of human macrophages. (D) Data from n = 3 healthy human macrophage donors in independent experiments are summarized, with each dot representing the Mtb burden for each individual patient with TB relative to control polyclonal IgG. The median and 95% confidence interval (CI) are shown. The dashed line shows the median of endemic IGRA− control individuals. The significance was determined by a Mann-Whitney U test.

Article Snippet: Mouse Anti-Human IgG2 Fc-PE HP6002 , SouthernBiotech , Cat#9070-09; RRID:AB_2796639.

Techniques: Derivative Assay, Infection, Bacteria, Control, MANN-WHITNEY

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: (A) The relationships between ESAT-6 and CFP-10 IgG levels and subclasses and intracellular Mtb burden within individuals with latent and active TB were evaluated by Spearman correlation. Heatmaps depict the Spearman rank correlation coefficient, ** p ≤ 0.01, and ^ stands for significance after adjustment for multiple comparisons by Benjamini-Hochberg. The scatterplot shows ESAT-6 and CFP-10 IgG1, and the Mtb burden is shown in the scatterplot, with each dot representing each individual with latent TB. (B) As a control, the relationships between Mtb cell-wall IgG levels and subclasses and intracellular Mtb burden are shown.

Article Snippet: Mouse Anti-Human IgG2 Fc-PE HP6002 , SouthernBiotech , Cat#9070-09; RRID:AB_2796639.

Techniques: Control

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: (A) Each column in the histogram depicts the intracellular Mtb burden of one individual patient with latent (light gray) and active (dark gray) TB as in . The dashed line represents the intracellular Mtb burden with control IgG. (A and B) The anti- Mtb activity of IgG from each individual patient with TB was determined by the difference in Mtb burden between control and patient IgG (red). (C) For the n = 17 latent and n = 14 active TB samples with detectable anti- Mtb activities relative to ESAT-6 and CFP-10 IgG1, the relationships to ESAT-6 and CFP-10 IgG Fc glycans as determined by Spearman correlations are listed with ^ marking the significance after adjustment for multiple comparisons by Benjamini-Hochberg. (D) N-glycans from anti-ESAT-6 and CFP-10 mAb were enzymatically removed with PNGase F and then used to treat Mtb -infected primary human monocyte-derived macrophages. The intracellular Mtb burden is shown relative to a no-antibody (Ab) control. The graph summarizes data for n = 6 healthy macrophage donors, with each line representing a single donor. (E) An L234A and L235A (LALA) variant of anti-ESAT-6 and CFP-10 mAb was used to treat Mtb -infected primary human monocyte-derived macrophages. Each line represents a single healthy macrophage donor ( n = 9). The significance was determined by a Wilcoxon matched-pairs signed rank test.

Article Snippet: Mouse Anti-Human IgG2 Fc-PE HP6002 , SouthernBiotech , Cat#9070-09; RRID:AB_2796639.

Techniques: Control, Activity Assay, Infection, Derivative Assay, Variant 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: 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: 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: Mouse anti-human IgG2 Fc-BIOT (clone HP6002); beads loaded at 25 nM , SouthernBiotech , Cat#9070-08; RRID: AB_2796638.

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: Mouse anti-human IgG2 Fc-BIOT (clone HP6002); beads loaded at 25 nM , SouthernBiotech , Cat#9070-08; RRID: AB_2796638.

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: Mouse anti-human IgG2 Fc-BIOT (clone HP6002); beads loaded at 25 nM , SouthernBiotech , Cat#9070-08; RRID: AB_2796638.

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: Mouse anti-human IgG2 Fc-BIOT (clone HP6002); beads loaded at 25 nM , SouthernBiotech , Cat#9070-08; RRID: AB_2796638.

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: Mouse anti-human IgG2 Fc-BIOT (clone HP6002); beads loaded at 25 nM , SouthernBiotech , Cat#9070-08; RRID: AB_2796638.

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