mouse α human igg3 hinge  (SouthernBiotech)

Journal: Journal of Human Immunity

Article Title: Oncostatin M receptor deficiency as a novel candidate genetic cause of autosomal recessive hyper-IgE syndrome

doi: 10.70962/jhi.20250119

Figure Lengend Snippet: B cell−extrinsic elevation of circulating plasmablasts and increase in IgE isotype switching in the patient. (A) Flow plots of main B cell subsets (top row) and antigen-experienced B cell subsets (bottom row) in one healthy control (left panels) and the patient (right panels). Representative of two independent analyses. (Data shown in rows 1 and 2, column 1 are identical to data in , row 1, column 4 and 5.) (B) Gate frequencies for antigen-experienced B cell subsets of five healthy controls (HCs) and the patient. Representative of two independent analyses. (C) Fraction of IgE class-switched cells among antigen-experienced B cell subsets of three healthy controls and the patient. The individual points for healthy control and patient represent one each of three sampling methods (conventional Ficoll prep, separator tubes, and whole-blood RBC lysis) from one experiment. (D) Schematic overview of iGB culture setup (created with BioRender). (E) Cell counts for one representative of two similar experiments. (F) IgM, IgG1, IgG3, and IgE secretion in cultures, normalized to cell count. Full gating strategies are shown in . Statistics were calculated using the unpaired t test. * = P < 0.05. aMBC_PC, aMBC/plasma cell; DN, double-negative; MBC_only, conventional memory B cell; SWmem, switched memory; T1, T2, T3, transitional 1, 2, 3 subsets.

Article Snippet: Wells of FluoroNunc MaxiSorp 96-well plates were coated with 100 μl capture antibody diluted in PBS, either 0.5 μg/ml goat α-human IgM-Hc (2023-01; Southern Biotech), 5 μg/ml mouse α-human IgG1-Fc (9054-01; Southern Biotech), 1 μg/ml mouse α-human IgG3-hinge (9210-01; Southern Biotech), or 2.5 μg/ml mouse α-human IgE-Fc (9240-01; Southern Biotech).

Techniques: Control, Sampling, Red Blood Cell Lysis, Cell Characterization, Clinical Proteomics

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Experimental timeline of the immunisation (i.e., vaccination) against OVA and subsequent tumour inoculation of the OVA-expressing B16F10 cell lines in mice. B) Titers of anti-OVA IgG per IgG subtypes in the plasma of the mice at d18. C) Percentage of OVA-specific CD8 + T cells against the immunogenic epitope OVA 257-264 (SIINFEKL) in the blood at d18 in pre-immunised mice (vax) or naïve mice (no vax) (n ≥ 3, mean ± SD, unpaired t-test). D, E) B16F10 melanoma cells, either wild-type (WT) or genetically modified to express high ( HI ) or low ( LO ) doses of membrane-bound OVA (B16mOVA) or soluble OVA (B16-OVA), were injected intradermally in C57BL/6 mice. Tumour growth (D) and associated survival (E) of the different OVA-expressing B16 cell lines in pre-immunised mice (n ≥ 5, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d10, d14 or d26 for tumour growth, log-rank tests for survival). F) Potential anti-tumour mechanisms engaged by soluble (left) or membrane-bound (right) xenoantigens when expressed by cancer cells.

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Expressing, Clinical Proteomics, Genetically Modified, Membrane, Injection

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Immunofluorescent staining of B16F10 WT melanoma with the TRP1-targeting TA99 antibody (scale bar = 100 μm). B) B16F10 WT tumour growth upon treatment with TA99 or IgG2a isotype control (n ≥ 4, mean ± SEM, Mann-Whitney test on d11). C) Design of FabTRP-OVA fusion protein and analysis of its production by SDS-PAGE (expected size = 92,3 kDa). D) Illustration of the FabTRP-OVA fusion protein approach for targeting the xenoantigen to the cancer cell membrane. E) Representative flow cytometry plot of FabTRP-OVA, OVA and no OVA (i.e., secondary antibody only) binding to the surface of B16F10 WT cells. F) B16F10 WT tumour growth when treated with FabTRP-OVA in pre-immunised or naïve mice (n ≥ 4, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d12). G) B16F10 WT tumour growth when treated with FabTRP-OVA in combination with anti-PD-1 checkpoint blocade therapy (n ≥ 4, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d12). H) Experimental treatment timeline of xenoantigen delivery (i.e., FabTRP-OVA, OVA or PBS (vehicle only)) and anti-PD-1 checkpoint blockade therapy in B16F10 WT in mice pre-immunised against OVA (Vax) or naïve (No vax). I, J) B16F10 WT tumour growth (I) and associated mouse survival (J) upon treatment with FabTRP-OVA, OVA or PBS in combination with anti-PD-1 checkpoint blockade therapy (n ≥ 5, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d16, log-rank tests for survival).

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Staining, Control, MANN-WHITNEY, SDS Page, Membrane, Flow Cytometry, Binding Assay

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Experimental timeline of the immunisation (i.e., vaccination) against varicella VZVO using Varivax or a combination of gE/CpG. B) IFΝγ quantification in the supernatant of gE restimulated (+) or non-restimulated (-) splenocytes, collected from mice immunized with Varivax, gE/CpG or PBS (i.e., naïve). Ionomycin+PMA was used as a positive control. C) Titers of anti-gE total IgG and per IgG subtypes in the plasma of the pre-immunised mice at d55 (before tumour implantation). D) Experimental treatment timeline of xenoantigen delivery (i.e., Varivax, gE or PBS (vehicle only)) and anti-PD-1 checkpoint blockade treatment in B16F10 WT in mice pre-immunised with Varivax against varicella VZVO . E, F) B16F10 WT tumour growth (E) and associated mouse survival (F) upon treatment with Varivax, gE or PBS in combination with anti-PD-1 checkpoint blockade (n ≥ 7, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d16, log-rank tests for survival).

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Positive Control, Clinical Proteomics

Journal: The Journal of Infectious Diseases

Article Title: Serum IgG1 and IgG3 Antibodies to Chlamydia trachomatis Pgp3 and Hsp60 in Men of Subfertile Couples

doi: 10.1093/infdis/jiaf535

Figure Lengend Snippet: Overlap between Chlamydia trachomatis Pgp3 immunoglobulin G1 (IgG1), Pgp3 immunoglobulin G3 (IgG3), Hsp60 IgG1, and Hsp60 IgG3 seropositivity.

Article Snippet: To test for IgG3 antibodies, Mouse Anti-Human IgG3 Hinge-AP (HP6050) (Southern Biotech) was used.

Techniques:

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