cd25-pe clone: bc96 antibody  (Thermo Fisher)

Journal: iScience

Article Title: Exploring the dynamics and influencing factors of CD4 T cell activation using single-cell RNA-seq

doi: 10.1016/j.isci.2023.107588

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Article Snippet: CD25-PE (clone: BC96) , eBioscience , Cat #12-0259-42; RRID: AB_1659682.

Techniques: Recombinant, SYBR Green Assay, Real-time Polymerase Chain Reaction, Software

Journal: Frontiers in Immunology

Article Title: B cells from anti-thyroid antibody positive, infertile women show hyper-reactivity to BCR stimulation

doi: 10.3389/fimmu.2022.1039166

Figure Lengend Snippet: Different naive and memory B cell ratio in levothyroxine treated and ATA+, euthyroid, infertile patients Prevalence of naive and memory B cell subsets (A) , B cell memory subpopulations (B) and CD25 + B cell subpopulations (C) HC: healthy controls (n=12); HT: Hashimoto’s thyroiditis, H1: hypothyroid patients with HT before treatment (n=14); H2: euthyroid patients with HT, after levothyroxine-treatment, (n=12); HIE: HT, infertile, euthyroid patients (n=14); NSw: non-switched; Sw: switched, DN: double negative memory B cells. Differences between the groups and within the B cell subpopulations were assessed with two-way ANOVA and Tukey’s post hoc test except H1-H2 comparisons where a mixed-effects analysis and the Sidak’s multiple comparisons tests were used because these groups represent the same individuals in a repeated sampling before and during treatment. Bars represent the mean ± standard deviation (SD) of percentages of lymphocyte subpopulation in each group. *p < 0.05, **p < 0.01.

Article Snippet: PBMCs (8 x 10 6 cells) were stained with the appropriate combination of fluorochrome-conjugated anti-human antibodies in 100 μl of media in order to differentiate B lymphocyte subsets: CD5 PerCP (clone UCHT2), CD19 PE (clone SJ25C1), IgD APC (clone IA6-2), CD25 BV421 (clone BC96) (all from Sony Biotechnology), CD27 PE-Cy7(clone M-T271, BioLegend).

Techniques: Sampling, Standard Deviation

Journal: Frontiers in Immunology

Article Title: B cells from anti-thyroid antibody positive, infertile women show hyper-reactivity to BCR stimulation

doi: 10.3389/fimmu.2022.1039166

Figure Lengend Snippet: Basal calcium level is elevated in B cells from ATA+, euthyroid, infertile patients Baseline median fluorescence intensity of Fluo4 in B cell subpopulations (A) , CD25- (C) and CD25+ (E) B cells. Cross tabulation shows the p values when comparing patient groups without the subpopulation-based grouping of B cells (B) , CD25- (D) and CD25+ cells (F) . HC: healthy controls (n=12); HT: Hashimoto’s thyroiditis, H1: hypothyroid patients with HT before treatment (n=14); H2: euthyroid patients with HT, after levothyroxine-treatment, (n=12); HIE: HT, infertile, euthyroid patients (n=14); NSw: non-switched; Sw: switched, DN: double negative memory B cells. Differences between the groups and within the B cell subpopulations were assessed with two-way ANOVA and Tukey’s post hoc test except H1-H2 comparisons, where a mixed-effects analysis and the Sidak’s multiple comparisons test were used because these groups represent the same individuals in a repeated sampling before and during treatment. Data are presented as follows: middle line represents the median, dot represents the mean, box: interquartile range, whiskers: min-max. *p < 0.05, **p < 0.01; Significant p values are red, non-significant p values are written in blue.

Article Snippet: PBMCs (8 x 10 6 cells) were stained with the appropriate combination of fluorochrome-conjugated anti-human antibodies in 100 μl of media in order to differentiate B lymphocyte subsets: CD5 PerCP (clone UCHT2), CD19 PE (clone SJ25C1), IgD APC (clone IA6-2), CD25 BV421 (clone BC96) (all from Sony Biotechnology), CD27 PE-Cy7(clone M-T271, BioLegend).

Techniques: Fluorescence, Sampling

Journal: Frontiers in Immunology

Article Title: B cells from anti-thyroid antibody positive, infertile women show hyper-reactivity to BCR stimulation

doi: 10.3389/fimmu.2022.1039166

Figure Lengend Snippet: Naive B cells show higher Ca 2+ signal upon BCR stimulation in ATA+ euthyroid, infertile patients Calcium flux kinetic data of naive B, naive CD25- and naive CD25+ B cells (A) maximum value, (B) ending value, (C) time to reach maximum, (D) time to first 50% value, (E) time from first 50% to maximum, (F) time from maximum to second 50% value, (G) slope at the first 50% value, (H) slope at second 50% value, (I) area under curve (AUC) value. HC: healthy controls (n=12); HT: Hashimoto’s thyroiditis, H1: hypothyroid patients with HT before treatment (n=14); H2: euthyroid patients with HT, after levothyroxine-treatment, (n=12); HIE: HT, infertile, euthyroid patients (n=14); NSw: non-switched; Sw: switched, DN: double negative memory B cells. The kinetic data were analyzed by the FACSKin software. Differences between the groups were assessed with two-way ANOVA and Tukey’s post hoc test except H1-H2 comparisons where a mixed-effects analysis and the Tukey’s multiple comparisons test were used because these groups represent the same individuals in a repeated sampling before and during treatment. Data are depicted as individual values, middle line represents the mean, whiskers are set to standard deviation (SD) *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: PBMCs (8 x 10 6 cells) were stained with the appropriate combination of fluorochrome-conjugated anti-human antibodies in 100 μl of media in order to differentiate B lymphocyte subsets: CD5 PerCP (clone UCHT2), CD19 PE (clone SJ25C1), IgD APC (clone IA6-2), CD25 BV421 (clone BC96) (all from Sony Biotechnology), CD27 PE-Cy7(clone M-T271, BioLegend).

Techniques: Software, Sampling, Standard Deviation

Journal: Frontiers in Immunology

Article Title: B cells from anti-thyroid antibody positive, infertile women show hyper-reactivity to BCR stimulation

doi: 10.3389/fimmu.2022.1039166

Figure Lengend Snippet: Memory B cells of ATA+, euthyroid, infertile patients have increased calcium flux upon BCR activation Selected calcium flux kinetic data of memory B cell subsets: Non-switched (NSw) memory B cells, CD25- and CD25+ NSw cells: (A) maximum value, (B) ending value, (C) area under curve (AUC) value. Immunoglobulin class-switched (Sw) memory B cells, CD25- and CD25+ Sw cells: (D) maximum value, (E) ending value, (F) area under curve (AUC) value. CD27-, IgD- double-negative (DN) memory B cells, CD25- and CD25+ DN cells: (G) maximum value, (H) ending value, (I) area under curve (AUC) value. HC: healthy controls (n=12); HT: Hashimoto’s thyroiditis, H1: hypothyroid patients with HT before treatment (n=14); H2: euthyroid patients with HT, after levothyroxine-treatment, (n=12); HIE: HT, infertile, euthyroid patients (n=14). The kinetic data were analyzed by the FACSKin software. Differences between the groups were assessed with two-way ANOVA and Tukey’s post hoc test except H1-H2 comparisons where a mixed-effects analysis and the Tukey’s multiple comparisons test were used because these groups represent the same individuals in a repeated sampling before and during treatment. Data are depicted as individual values, middle line represents the mean, whiskers are set to standard deviation (SD). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: PBMCs (8 x 10 6 cells) were stained with the appropriate combination of fluorochrome-conjugated anti-human antibodies in 100 μl of media in order to differentiate B lymphocyte subsets: CD5 PerCP (clone UCHT2), CD19 PE (clone SJ25C1), IgD APC (clone IA6-2), CD25 BV421 (clone BC96) (all from Sony Biotechnology), CD27 PE-Cy7(clone M-T271, BioLegend).

Techniques: Activation Assay, Software, Sampling, Standard Deviation

Journal: Antioxidants

Article Title: Role of Oxidative Stress in Vascular Low-Grade Inflammation Initiation Due to Acute Salt Loading in Young Healthy Individuals

doi: 10.3390/antiox11030444

Figure Lengend Snippet: Gating strategy for the assessment of peripheral blood regulatory T cells (Treg) by flow cytometry. Panel ( A ) shows representative dot plots illustrating gating strategy, including exclusion of doublets using forward scatter area (FSC-A) versus forward scatter width (FSC-W) analysis ( A - i ), gating on live cells negative for aminereactive fixable viability dye ( A - ii ), lymphocytes ( A - iii ), and CD3 + T cells ( A - iv ). Next, total CD25 and Foxp3, expressing T cells among CD4 + CD127 low population, were analyzed (Panel ( B )). ( B - i ) shows representative gating strategy, ( B - ii ) representative samples from LS and HS protocol. FlowLogic software was used for data analysis and illustration.

Article Snippet: In order to determine expression of Treg specific cell surface antigens, the following antibody mixture was used: CD3 FITC (clone: OKT3, eBioscienceTM, Affymetrix by Thermo Fisher Scientific, CA, USA), CD4 PerCP-eFluorTM 710 (clone: SK3, eBioscienceTM), CD127 PE-Cy7 (clone: eBioRDR5, eBioscienceTM), and CD25 APC (clone: BC96, eBioscienceTM); the Foxp3 PE (clone: 235A/E7, eBioscienceTM) antibody was used for the detection of Treg signature transcription factor Foxp3.

Techniques: Flow Cytometry, Expressing, Software

Journal: Arthritis Research & Therapy

Article Title: Characterizing memory T helper cells in patients with psoriasis, subclinical, or early psoriatic arthritis using a machine learning algorithm

doi: 10.1186/s13075-021-02714-5

Figure Lengend Snippet: Unbiased clustering of memory T helper cells using FlowSOM algorithm. PBMCs of psoriasis and PsA patients and healthy controls were isolated and used for Flow cytometry. Doublets and dead cells were excluded. A Living CD3 + CD4 + CD45RO + CD25 low/int cells were gated for each subject and exported, arcsinh-transformed, down-sampled to 6000 cells per sample, and aggregated per group. The 3 aggregated files were used to generate a FlowSOM tree and cells were clustered into 100 nodes. The nodes were clustered in 12 metaclusters with hierarchical clustering, indicated by the background colors. B Table of metaclusters and the known labels for the different memory T helper subsets. C Median expression of the 6 individual surface markers (CCR6, CD25, CCR4, CXCR3, CCR10, and CLA) plotted in the generated FlowSOM tree

Article Snippet: Or cells were incubated with the second panel including CD3 (clone UCHT1; BD Biosciences), CD4 (clone SK3; BD Biosciences), CD25 (clone bc96; Sony), CD45RO (clone UCHL1; eBioscience), CD127 (clone hIL-7R-M21;BD Biosciences), and CLA (clone HECA-452; BD Biosciences).

Techniques: Isolation, Flow Cytometry, Transformation Assay, Expressing, Generated