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untouched memory cd4 t cell isolation kit  (Miltenyi Biotec)


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    Miltenyi Biotec untouched memory cd4 t cell isolation kit
    (A) Diagram depicting the culture model used to generate time course samples from healthy human <t>CD4+</t> memory T cells. Cells were stimulated with CD3/CD28 Dynabeads for 12 days, with fresh beads plus full media change every other day (fresh media alone added on alternate days). Samples were collected on days highlighted in bold. (B) Multiomics workflow (see for details). Experiments were performed in biological triplicate, with proteomics, phosphoproteomics, and metabolomics done on same samples. (C) ELISA-based quantification of secreted cytokines (IL-2, IFN-γ, TNF-α) on days 0, 6, 10, and 12, 3 replicates per time point. (D) Cell counts were taken each time point, and average cell division compared to previous time point calculated. Cell viability remained stable throughout at ~75%–85%. (E) Flow cytometric analysis of inhibitory receptors PD-1, TIM-3, and TIGIT at resting, day 6, and day 12 time points. Data normalized to mode for accurate population comparisons to account for differences in number of cells analyzed. The data underlying can be found in the supplemental flow cytometry files uploaded to flowrepository.org . Figure created with BioRender.com .
    Untouched Memory Cd4 T Cell Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 95/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+phosphorylated+rtk+array+kit/Memory+CD4%2B+T+Cell+Isolation+Kit%2C+human/pmc11703073-398-22-28
    Average 95 stars, based on 36 article reviews
    untouched memory cd4 t cell isolation kit - by Bioz Stars, 2026-11
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    Images

    1) Product Images from "Multiomic profiling of chronically activated CD4+ T cells identifies drivers of exhaustion and metabolic reprogramming"

    Article Title: Multiomic profiling of chronically activated CD4+ T cells identifies drivers of exhaustion and metabolic reprogramming

    Journal: PLOS Biology

    doi: 10.1371/journal.pbio.3002943

    (A) Diagram depicting the culture model used to generate time course samples from healthy human CD4+ memory T cells. Cells were stimulated with CD3/CD28 Dynabeads for 12 days, with fresh beads plus full media change every other day (fresh media alone added on alternate days). Samples were collected on days highlighted in bold. (B) Multiomics workflow (see for details). Experiments were performed in biological triplicate, with proteomics, phosphoproteomics, and metabolomics done on same samples. (C) ELISA-based quantification of secreted cytokines (IL-2, IFN-γ, TNF-α) on days 0, 6, 10, and 12, 3 replicates per time point. (D) Cell counts were taken each time point, and average cell division compared to previous time point calculated. Cell viability remained stable throughout at ~75%–85%. (E) Flow cytometric analysis of inhibitory receptors PD-1, TIM-3, and TIGIT at resting, day 6, and day 12 time points. Data normalized to mode for accurate population comparisons to account for differences in number of cells analyzed. The data underlying can be found in the supplemental flow cytometry files uploaded to flowrepository.org . Figure created with BioRender.com .
    Figure Legend Snippet: (A) Diagram depicting the culture model used to generate time course samples from healthy human CD4+ memory T cells. Cells were stimulated with CD3/CD28 Dynabeads for 12 days, with fresh beads plus full media change every other day (fresh media alone added on alternate days). Samples were collected on days highlighted in bold. (B) Multiomics workflow (see for details). Experiments were performed in biological triplicate, with proteomics, phosphoproteomics, and metabolomics done on same samples. (C) ELISA-based quantification of secreted cytokines (IL-2, IFN-γ, TNF-α) on days 0, 6, 10, and 12, 3 replicates per time point. (D) Cell counts were taken each time point, and average cell division compared to previous time point calculated. Cell viability remained stable throughout at ~75%–85%. (E) Flow cytometric analysis of inhibitory receptors PD-1, TIM-3, and TIGIT at resting, day 6, and day 12 time points. Data normalized to mode for accurate population comparisons to account for differences in number of cells analyzed. The data underlying can be found in the supplemental flow cytometry files uploaded to flowrepository.org . Figure created with BioRender.com .

    Techniques Used: Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Flow Cytometry

    Mean fluorescence intensity of PD-1 in CD4+ T cells from both HIV+ and HIV- patients (grouped due to observing same effect in both) for CD4+ cells that are CD276+ or FLT-1+. The data underlying this figure can be found in the supplemental flow cytometry files uploaded to flowrepository.org . Figure created with BioRender.com .
    Figure Legend Snippet: Mean fluorescence intensity of PD-1 in CD4+ T cells from both HIV+ and HIV- patients (grouped due to observing same effect in both) for CD4+ cells that are CD276+ or FLT-1+. The data underlying this figure can be found in the supplemental flow cytometry files uploaded to flowrepository.org . Figure created with BioRender.com .

    Techniques Used: Fluorescence, Flow Cytometry

    (A) Causal network analysis leveraging annotated interactions (transcription factor-target, kinase-substrate, phosphatase-substrate) as a framework to evaluate protein expression changes (late exhaustion (day 12) versus resting, taking into account previous time point comparisons) to predict key nodes (regulators) impacting most changes seen across the data set. (B) Pruned version of a large subnetwork with p300 (black outline) at main hub. Orange circles mark proteins of interest mentioned in main text. (C) Depiction of experimental design of p300 inhibition studies. (D) Flow cytometric analysis of PD-1 and TIM-3 showing shifts in fluorescent intensity of cell populations in treated versus untreated (cell number normalized to mode). Cell numbers shown for each plot are 20,312 for p300 treated and 30,438 for DMSO, which corresponds to all CD4+ T cells from the sample (excluding doublets and debris, see for gating), corresponding to >95% of sample. (E) Bar chart of mean fluorescent intensity showing significant changes ( T test) in PD-1 and TIM-3 during p300 inhibition versus controls (antigen removed for 2 days, or treatment with T-cell activation inhibitor Dasatinib); *, p -value <0.05, ***, p -value <0.005. (F) Heatmap showing recruitment of cofactors (p300, class I HDACs, KAT2B) to transcription factor family DNA motifs as measured using CASCADE protein-binding microarrays. The data underlying this figure can be found in and Data (A, B), supplemental flow cytometry files uploaded to flowrepositroy.org (D), and the (E, F). Figure created with BioRender.com .
    Figure Legend Snippet: (A) Causal network analysis leveraging annotated interactions (transcription factor-target, kinase-substrate, phosphatase-substrate) as a framework to evaluate protein expression changes (late exhaustion (day 12) versus resting, taking into account previous time point comparisons) to predict key nodes (regulators) impacting most changes seen across the data set. (B) Pruned version of a large subnetwork with p300 (black outline) at main hub. Orange circles mark proteins of interest mentioned in main text. (C) Depiction of experimental design of p300 inhibition studies. (D) Flow cytometric analysis of PD-1 and TIM-3 showing shifts in fluorescent intensity of cell populations in treated versus untreated (cell number normalized to mode). Cell numbers shown for each plot are 20,312 for p300 treated and 30,438 for DMSO, which corresponds to all CD4+ T cells from the sample (excluding doublets and debris, see for gating), corresponding to >95% of sample. (E) Bar chart of mean fluorescent intensity showing significant changes ( T test) in PD-1 and TIM-3 during p300 inhibition versus controls (antigen removed for 2 days, or treatment with T-cell activation inhibitor Dasatinib); *, p -value <0.05, ***, p -value <0.005. (F) Heatmap showing recruitment of cofactors (p300, class I HDACs, KAT2B) to transcription factor family DNA motifs as measured using CASCADE protein-binding microarrays. The data underlying this figure can be found in and Data (A, B), supplemental flow cytometry files uploaded to flowrepositroy.org (D), and the (E, F). Figure created with BioRender.com .

    Techniques Used: Expressing, Inhibition, Activation Assay, Protein Binding, Flow Cytometry

    UMAPs of CD4+ subtypes and CD8+ T exhausted tumor lymphocytes showing gene expression of (A) exhaustion-associated genes, and (B) gene expression of proteins identified in the model presented by Lawton and colleagues. UMAPs made using Single Cell Portal, Tarhan and colleagues. The data from this figure and tool used to make it can be found at https://singlecell.broadinstitute.org/single_cell/study/SCP1039/a-single-cell-and-spatially-resolved-atlas-of-human-breast-cancers . Figure created with BioRender.com .
    Figure Legend Snippet: UMAPs of CD4+ subtypes and CD8+ T exhausted tumor lymphocytes showing gene expression of (A) exhaustion-associated genes, and (B) gene expression of proteins identified in the model presented by Lawton and colleagues. UMAPs made using Single Cell Portal, Tarhan and colleagues. The data from this figure and tool used to make it can be found at https://singlecell.broadinstitute.org/single_cell/study/SCP1039/a-single-cell-and-spatially-resolved-atlas-of-human-breast-cancers . Figure created with BioRender.com .

    Techniques Used: Gene Expression

    Related Articles

    Cell Isolation:

    Article Title: L-Phenylalanine is a metabolic checkpoint of human Th2 cells
    Article Snippet: CD4+T cell Isolation Kit, human , Miltenyi Biotec , Cat#130-096-533. .. Memory CD4+T cell Isolation Kit, human , Miltenyi Biotec , Cat130-091-893. .. Seahorse Glycolytic Rate Assay kit , Agilent Technologies , Cat#103344-100.

    Article Title: Post-COVID impairment of memory T cell responses to community-acquired pathogens can be rectified by activating cellular metabolism
    Article Snippet: .. CD4+CD45RO+ memory T cells were isolated by negative selection using the CD4 memory T cell isolation kit from Miltenyi Biotec. .. Cells were lysed and RNA extracted using the RNEasy Mini Kit (Qiagen).

    Article Title: Rationale for multi-epitope TGFβ vaccination in pancreatic cancer: evidence from immunologic and clinical correlates
    Article Snippet: Fc receptors were blocked for 10 min, followed by 30 min staining with HLA-DR,DP,DQ-FITC (BD Biosciences) and NIR viability dye. .. CD4+CD45RO+ memory T cells were enriched using the human Memory CD4+ T Cell Isolation Kit (Miltenyi Biotec). .. CD14+ monocytes were isolated from PBMCs using MACS CD14 MicroBeads and pulsed with peptide (10 nM) for 2 h. CD4+CD45RO+ memory T cells were isolated from the CD14 − fraction and co-cultured with peptide-pulsed monocytes.

    Article Title: Refined single-cell profiling captures a CCR5 high CD4 + cytotoxic T-cell precursor in multiple sclerosis.
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation with Ficoll–Paque (GE Healthcare). .. Memory CD4+ T cells were subsequently purified from PBMCs through magnetic-activated cell sorting (MACS) utilising the Memory CD4+ T Cell Isolation Kit (Miltenyi Biotec, Cat130-091-893) on the autoMACS Pro Separator (Miltenyi Biotec), following the manufacturer’s protocol. .. MACS isolated CD4+ memory cells were cryopreserved in RPMI 1640 medium (Gibco, Cat# 11875093) supplemented with 20% calf serum (FCS; Gibco, Cat# 10270-106) and 10% dimethyl sulfoxide (Sigma–Aldrich), and stored at −80 ◦C until further use in experiments.

    Article Title: Rationale for multi-epitope TGFβ vaccination in pancreatic cancer: evidence from immunologic and clinical correlates.
    Article Snippet: Fc receptors were blocked for 10 min, followed by 30min staining with HLA-DR,DP,DQ-FITC (BD Biosciences) and NIR viability dye. .. Isolation and identification of circulating memory CD4+ T cells specific to TGFβ-33 CD4+CD45RO+ memory T cells were enriched using the human Memory CD4+ T Cell Isolation Kit (Miltenyi Biotec).42 CD14+ monocytes were isolated from PBMCs using MACS CD14 MicroBeads16 and pulsed with peptide (10 nM) for 2 h. CD4+CD45RO+ memory T cells were isolated from the CD14− fraction and cocultured with peptide-pulsed monocytes. ..

    Article Title: L-Phenylalanine is a metabolic checkpoint of human Th2 cells
    Article Snippet: .. Next, total CD3 + CD4 + T cells or memory CD3 + CD4 + T cells were isolated using the human CD4 + T cell Isolation Kit or Memory CD4 + T cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany), for respective cell types, according to manufacturer instructions on the autoMACS Pro Separator (Miltenyi Biotec, Bergisch Gladbach, Germany). .. Isolated cells were then transferred to RPMI1640 medium (Lonza, Basel, Switzerland) with 10% heat inactivated FCS (Merck, Darmstadt, Germany), 1% NEA (Merck, Darmstadt, Germany) and 100U/mL rhIL-2 (PeproTech, Thermo Fisher Scientific, Waltham, MA, USA) (R10+IL2).

    Isolation:

    Article Title: Post-COVID impairment of memory T cell responses to community-acquired pathogens can be rectified by activating cellular metabolism
    Article Snippet: .. CD4+CD45RO+ memory T cells were isolated by negative selection using the CD4 memory T cell isolation kit from Miltenyi Biotec. .. Cells were lysed and RNA extracted using the RNEasy Mini Kit (Qiagen).

    Article Title: Rationale for multi-epitope TGFβ vaccination in pancreatic cancer: evidence from immunologic and clinical correlates.
    Article Snippet: Fc receptors were blocked for 10 min, followed by 30min staining with HLA-DR,DP,DQ-FITC (BD Biosciences) and NIR viability dye. .. Isolation and identification of circulating memory CD4+ T cells specific to TGFβ-33 CD4+CD45RO+ memory T cells were enriched using the human Memory CD4+ T Cell Isolation Kit (Miltenyi Biotec).42 CD14+ monocytes were isolated from PBMCs using MACS CD14 MicroBeads16 and pulsed with peptide (10 nM) for 2 h. CD4+CD45RO+ memory T cells were isolated from the CD14− fraction and cocultured with peptide-pulsed monocytes. ..

    Article Title: L-Phenylalanine is a metabolic checkpoint of human Th2 cells
    Article Snippet: .. Next, total CD3 + CD4 + T cells or memory CD3 + CD4 + T cells were isolated using the human CD4 + T cell Isolation Kit or Memory CD4 + T cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany), for respective cell types, according to manufacturer instructions on the autoMACS Pro Separator (Miltenyi Biotec, Bergisch Gladbach, Germany). .. Isolated cells were then transferred to RPMI1640 medium (Lonza, Basel, Switzerland) with 10% heat inactivated FCS (Merck, Darmstadt, Germany), 1% NEA (Merck, Darmstadt, Germany) and 100U/mL rhIL-2 (PeproTech, Thermo Fisher Scientific, Waltham, MA, USA) (R10+IL2).

    Selection:

    Article Title: Post-COVID impairment of memory T cell responses to community-acquired pathogens can be rectified by activating cellular metabolism
    Article Snippet: .. CD4+CD45RO+ memory T cells were isolated by negative selection using the CD4 memory T cell isolation kit from Miltenyi Biotec. .. Cells were lysed and RNA extracted using the RNEasy Mini Kit (Qiagen).

    other:

    Article Title: O-GlcNAcylation and low glycolysis underpin Th2 polarization by dendritic cells
    Article Snippet: Naïve CD4 T cells for assessment of DC capacity for Th1/2 polarization were isolated from allogenic buffy coat PBMCs using a naïve human CD4 T cell isolation kit (#480042, BioLegend).

    Purification:

    Article Title: Refined single-cell profiling captures a CCR5 high CD4 + cytotoxic T-cell precursor in multiple sclerosis.
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation with Ficoll–Paque (GE Healthcare). .. Memory CD4+ T cells were subsequently purified from PBMCs through magnetic-activated cell sorting (MACS) utilising the Memory CD4+ T Cell Isolation Kit (Miltenyi Biotec, Cat130-091-893) on the autoMACS Pro Separator (Miltenyi Biotec), following the manufacturer’s protocol. .. MACS isolated CD4+ memory cells were cryopreserved in RPMI 1640 medium (Gibco, Cat# 11875093) supplemented with 20% calf serum (FCS; Gibco, Cat# 10270-106) and 10% dimethyl sulfoxide (Sigma–Aldrich), and stored at −80 ◦C until further use in experiments.

    FACS:

    Article Title: Refined single-cell profiling captures a CCR5 high CD4 + cytotoxic T-cell precursor in multiple sclerosis.
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation with Ficoll–Paque (GE Healthcare). .. Memory CD4+ T cells were subsequently purified from PBMCs through magnetic-activated cell sorting (MACS) utilising the Memory CD4+ T Cell Isolation Kit (Miltenyi Biotec, Cat130-091-893) on the autoMACS Pro Separator (Miltenyi Biotec), following the manufacturer’s protocol. .. MACS isolated CD4+ memory cells were cryopreserved in RPMI 1640 medium (Gibco, Cat# 11875093) supplemented with 20% calf serum (FCS; Gibco, Cat# 10270-106) and 10% dimethyl sulfoxide (Sigma–Aldrich), and stored at −80 ◦C until further use in experiments.

    Magnetic Cell Separation:

    Article Title: Refined single-cell profiling captures a CCR5 high CD4 + cytotoxic T-cell precursor in multiple sclerosis.
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation with Ficoll–Paque (GE Healthcare). .. Memory CD4+ T cells were subsequently purified from PBMCs through magnetic-activated cell sorting (MACS) utilising the Memory CD4+ T Cell Isolation Kit (Miltenyi Biotec, Cat130-091-893) on the autoMACS Pro Separator (Miltenyi Biotec), following the manufacturer’s protocol. .. MACS isolated CD4+ memory cells were cryopreserved in RPMI 1640 medium (Gibco, Cat# 11875093) supplemented with 20% calf serum (FCS; Gibco, Cat# 10270-106) and 10% dimethyl sulfoxide (Sigma–Aldrich), and stored at −80 ◦C until further use in experiments.

    Article Title: Rationale for multi-epitope TGFβ vaccination in pancreatic cancer: evidence from immunologic and clinical correlates.
    Article Snippet: Fc receptors were blocked for 10 min, followed by 30min staining with HLA-DR,DP,DQ-FITC (BD Biosciences) and NIR viability dye. .. Isolation and identification of circulating memory CD4+ T cells specific to TGFβ-33 CD4+CD45RO+ memory T cells were enriched using the human Memory CD4+ T Cell Isolation Kit (Miltenyi Biotec).42 CD14+ monocytes were isolated from PBMCs using MACS CD14 MicroBeads16 and pulsed with peptide (10 nM) for 2 h. CD4+CD45RO+ memory T cells were isolated from the CD14− fraction and cocultured with peptide-pulsed monocytes. ..



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    Figure Lengend Snippet:

    Article Snippet: Human RTK Phosphorylation Antibody Array Kit , RayBiotech , Cat# AAH-PRTK-1-8.

    Techniques: Recombinant, Lysis, Electrophoresis, Cell Counting, Fluorescence, Staining, Phospho-proteomics, Ab Array, Plasmid Preparation, Software

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