antibodies igg4 mouse hp6025 bio rad cat  (Bio-Rad)

Journal: Journal of Translational Autoimmunity

Article Title: Anti-IgLON5 encephalitis is associated with anti-retinal immunological reactivity without retinal alteration

doi: 10.1016/j.jtauto.2026.100359

Figure Lengend Snippet: Indirect immunofluorescence on sections of monkey retina. The fluorescence observed is identified by a black arrow. (A) Patient 4 (IgG4); (B) Patient 1 (IgG4); (C) Patient 3 (IgG1); (D) Patient 5 (IgG4); (E) Patient 2 (IgG1); (F) Patient 1 (IgG4) after immunoadsorption of IgLON5 antibodies; (G) Control with macular edema (IgG1); (H) Control with anti-Hu encephalitis (IgG1); (I) Control with CAR syndrome (IgG4). The different layers of the retina are identified by their initials: pigment epithelium (pe), photoreceptor layer (pr), outer grain layer (og), outer plexiform layer (op), inner grain layer (ig), inner plexiform layer (ip), ganglion cell layer (gc), nerve fibre layer (nf).

Article Snippet: The reactivity of the patients’ sera and/or CSF against the retina was evaluated retrospectively with frozen samples (−80°, EXPLAINEUR biobank) through an indirect immunofluorescence technique using sections of monkey retina (Ref. FA1172-1005, Euroimmun), detected with an FITC-labelled secondary antibody anti-human IgAGM (Euroimmun conjugate) or directed against IgA (ref. F0204, DAKO), IgM (ref. F0203, DAKO), IgG1 (ref. 9052-02, Southern Biotech) and IgG4 (ref. 9200-02, Southern Biotech).

Techniques: Immunofluorescence, Fluorescence, Control

Journal: Transfusion

Article Title: IgG4 anti‐ CD47 : Erythrophagocytosis and potential transfusion complications

doi: 10.1111/trf.70111

Figure Lengend Snippet: IgG4 antibody induced erythrophagocytosis and enhancement of alloantibody phagocytosis when CD47 is blocked. (A) IgG4 anti‐K (KEL1) causes erythrophagocytosis which is enhanced when CD47 is blocked using a deglycosylated anti‐CD47. (B) Polyclonal anti‐D (WinRho) erythrophagocytosis is enhanced in a dose‐dependent manner with deglycosylated anti‐CD47 blocking antibody. Results are using Student's t ‐test, mean ± SD for triplicates, using GraphPad. * p < .05, ** p < .01, *** p < .001, **** p < .0001.

Article Snippet: Drug or patients' diluted plasma was used to opsonize blood group O RBCs, washed, and stained with a PE‐labeled mouse monoclonal anti‐human IgG4 Fc (Southern Biotech, clone HP6025, Cat. # 9200‐09) for 30 min at 4°C.

Techniques: Blocking Assay

Journal: Transfusion

Article Title: IgG4 anti‐ CD47 : Erythrophagocytosis and potential transfusion complications

doi: 10.1111/trf.70111

Figure Lengend Snippet: Schematic depicting that (A) IgG4 anti‐CD47 leads to robust, largely FcγRI‐dependent erythrophagocytosis, but also involvement of FcRIIa but not FcRIIIa; and (B) antibodies to CD47 with functional Fc regions may block CD47 but contribute to phagocytosis and show an additive phagocytosis with unrelated alloantibodies. Ani‐CD47 with a nonfunctional Fc region may block CD47‐SIRPα (blocking the “don't eat me” signal) and lead to significant erythrophagocytosis of unrelated red blood cell (RBC) alloantibodies and may augment delayed hemolytic transfusion reactions (DHTRs).

Article Snippet: Drug or patients' diluted plasma was used to opsonize blood group O RBCs, washed, and stained with a PE‐labeled mouse monoclonal anti‐human IgG4 Fc (Southern Biotech, clone HP6025, Cat. # 9200‐09) for 30 min at 4°C.

Techniques: Functional Assay, Blocking Assay

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