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mouse naïve cd4 t cell isolation kit  (Miltenyi Biotec)


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    Structured Review

    Miltenyi Biotec mouse naïve cd4 t cell isolation kit
    GPCR68 as a pH-Sensing regulator in T Cells and generation of GPCR68 fl/fl <t>CD4</t> Cre mice. (A) Schematic diagram of the effect of pH on T cell GPCR68 as well as tumor. (B) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with varying pH. RT-qPCR was performed to determine the expression of GPCR68 at various pH. (C) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 under different pH conditions, and GPCR68 protein expression was assessed by Western blot analysis. (D) To generate conditional knockout (CKO) of GPCR68 in T cells, GPCR68 fl/fl mice were crossed with CD4 Cre mice and generated GPCR68 fl/fl CD4 Cre (CKO). (E) Flow cytometry was used to determine the population of CD4 and CD8 cells in the lymph nodes (LN), thymus (THY), and spleen (SP) at the basal level in CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (F) Flow cytometry was used to determine the population of Foxp3+ Treg cells in the lymph nodes, thymus, and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (G-H) The population of F4/80+, CD11c+ (G), and B220+ (H) cells was determined in the lymph nodes and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (I-J) Flow cytometry was used to evaluate the CD4 + or CD8 + T cells for the determination of intracellular cytokines IFN-γ+ (I), or TNF-α+ (J) from the spleen and lymph nodes at basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. Student t-test was performed for comparison between the two groups. Data are mean ± SEM (n = 5), ∗ p < 0.05.
    Mouse Naïve Cd4 T Cell Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 438 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/isolation+kit/Naive+CD4%2B+T+Cell+Isolation+Kit%2C+mouse/pmc12963920-412-26-35
    Average 97 stars, based on 438 article reviews
    mouse naïve cd4 t cell isolation kit - by Bioz Stars, 2026-10
    97/100 stars

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    1) Product Images from "pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity"

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

    Journal: Bioactive Materials

    doi: 10.1016/j.bioactmat.2026.02.039

    GPCR68 as a pH-Sensing regulator in T Cells and generation of GPCR68 fl/fl CD4 Cre mice. (A) Schematic diagram of the effect of pH on T cell GPCR68 as well as tumor. (B) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with varying pH. RT-qPCR was performed to determine the expression of GPCR68 at various pH. (C) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 under different pH conditions, and GPCR68 protein expression was assessed by Western blot analysis. (D) To generate conditional knockout (CKO) of GPCR68 in T cells, GPCR68 fl/fl mice were crossed with CD4 Cre mice and generated GPCR68 fl/fl CD4 Cre (CKO). (E) Flow cytometry was used to determine the population of CD4 and CD8 cells in the lymph nodes (LN), thymus (THY), and spleen (SP) at the basal level in CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (F) Flow cytometry was used to determine the population of Foxp3+ Treg cells in the lymph nodes, thymus, and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (G-H) The population of F4/80+, CD11c+ (G), and B220+ (H) cells was determined in the lymph nodes and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (I-J) Flow cytometry was used to evaluate the CD4 + or CD8 + T cells for the determination of intracellular cytokines IFN-γ+ (I), or TNF-α+ (J) from the spleen and lymph nodes at basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. Student t-test was performed for comparison between the two groups. Data are mean ± SEM (n = 5), ∗ p < 0.05.
    Figure Legend Snippet: GPCR68 as a pH-Sensing regulator in T Cells and generation of GPCR68 fl/fl CD4 Cre mice. (A) Schematic diagram of the effect of pH on T cell GPCR68 as well as tumor. (B) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with varying pH. RT-qPCR was performed to determine the expression of GPCR68 at various pH. (C) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 under different pH conditions, and GPCR68 protein expression was assessed by Western blot analysis. (D) To generate conditional knockout (CKO) of GPCR68 in T cells, GPCR68 fl/fl mice were crossed with CD4 Cre mice and generated GPCR68 fl/fl CD4 Cre (CKO). (E) Flow cytometry was used to determine the population of CD4 and CD8 cells in the lymph nodes (LN), thymus (THY), and spleen (SP) at the basal level in CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (F) Flow cytometry was used to determine the population of Foxp3+ Treg cells in the lymph nodes, thymus, and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (G-H) The population of F4/80+, CD11c+ (G), and B220+ (H) cells was determined in the lymph nodes and spleen at the basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (I-J) Flow cytometry was used to evaluate the CD4 + or CD8 + T cells for the determination of intracellular cytokines IFN-γ+ (I), or TNF-α+ (J) from the spleen and lymph nodes at basal level in the CD4 Cre or GPCR68 fl/fl CD4 Cre mice. Student t-test was performed for comparison between the two groups. Data are mean ± SEM (n = 5), ∗ p < 0.05.

    Techniques Used: Isolation, Quantitative RT-PCR, Expressing, Western Blot, Knock-Out, Generated, Flow Cytometry, Comparison

    GPCR68 fl/fl CD4 Cre mice exhibit improved anti-tumor mmune responses. (A-C) Naïve CD4 + T cells were isolated from CD4 Cre or GPCR68 fl/fl CD4 Cre mice and activated using anti-CD3 and anti-CD28 using the culture media under physiologic neutral pH (7.4) or varying pH 6.0, 6.5, or 7.8. Flow cytometry plots showing the expression of IFN-γ and IL-2 in CD4 + T cells from CD4 Cre and GPCR68 fl/fl CD4 Cre mice. Each panel represents the frequency of IFN-γ + and IL-2 + cells. (B) Bar graph summarizing the percentage of IFN-γ + CD4 + T cells at each pH level for CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (C) Bar graph showing the percentage of IL-2 + CD4 + T cells at each pH for CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (D) Experimental timeline depicting tumor induction and treatment protocol in CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (E) Tumor growth curves in CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (F) Tumor weight in CD4 Cre versus GPCR68 fl/fl CD4 Cre mice at the time of harvesting on day 21. (G) Representative images of excised tumors at day 21. (H) Flow cytometric analysis of IFN-γ production by tumor-infiltrating CD4 + and CD8 + T cells. (I) Flow cytometric analysis of TNF-α production by tumor-infiltrating CD4 + and CD8 + T cells. Student t-test was performed for comparison between the two groups. Two-way ANOVA was used for multiple comparisons. Data are mean ± SEM (n = 5). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns = not significant.
    Figure Legend Snippet: GPCR68 fl/fl CD4 Cre mice exhibit improved anti-tumor mmune responses. (A-C) Naïve CD4 + T cells were isolated from CD4 Cre or GPCR68 fl/fl CD4 Cre mice and activated using anti-CD3 and anti-CD28 using the culture media under physiologic neutral pH (7.4) or varying pH 6.0, 6.5, or 7.8. Flow cytometry plots showing the expression of IFN-γ and IL-2 in CD4 + T cells from CD4 Cre and GPCR68 fl/fl CD4 Cre mice. Each panel represents the frequency of IFN-γ + and IL-2 + cells. (B) Bar graph summarizing the percentage of IFN-γ + CD4 + T cells at each pH level for CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (C) Bar graph showing the percentage of IL-2 + CD4 + T cells at each pH for CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (D) Experimental timeline depicting tumor induction and treatment protocol in CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (E) Tumor growth curves in CD4 Cre and GPCR68 fl/fl CD4 Cre mice. (F) Tumor weight in CD4 Cre versus GPCR68 fl/fl CD4 Cre mice at the time of harvesting on day 21. (G) Representative images of excised tumors at day 21. (H) Flow cytometric analysis of IFN-γ production by tumor-infiltrating CD4 + and CD8 + T cells. (I) Flow cytometric analysis of TNF-α production by tumor-infiltrating CD4 + and CD8 + T cells. Student t-test was performed for comparison between the two groups. Two-way ANOVA was used for multiple comparisons. Data are mean ± SEM (n = 5). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns = not significant.

    Techniques Used: Isolation, Flow Cytometry, Expressing, Comparison

    Physicochemical properties of BOLT, and BOLT reduces the growth of tumor cells. (A) Schematic of surface double-layer formation and ion release. (B) Negative zeta potential (−1.365 mV) and high conductivity (1.334 mS/cm), confirming colloidal stability and ion release. (C) Uniform particle size (∼1478 nm) across batches. (D) Interfacial pH buffering in PBS. (E) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with 6.0 pH and treated with various doses of BOLT. RT-qPCR was performed to determine the expression of Gpcr68 at various BOLT doses in activated T cells at acidic pH. (F) Anti-CD3 and anti-CD28 activated CD4 + T cells were treated with different doses of BOLT to determine the protein expression of GPCR68 using Western blot. (G-J) CCK8 assay was performed to analyze the effect of various pH on B16, MC38, 143B, and MG63 cell proliferation. (K-L) Effect of various doses of BOLT on the B16 and K7M2 cell growth to determine the IC-50 of BOLT. Error bars represent mean ± SEM. ∗∗ p < 0.01 and ∗ p < 0.05.
    Figure Legend Snippet: Physicochemical properties of BOLT, and BOLT reduces the growth of tumor cells. (A) Schematic of surface double-layer formation and ion release. (B) Negative zeta potential (−1.365 mV) and high conductivity (1.334 mS/cm), confirming colloidal stability and ion release. (C) Uniform particle size (∼1478 nm) across batches. (D) Interfacial pH buffering in PBS. (E) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with 6.0 pH and treated with various doses of BOLT. RT-qPCR was performed to determine the expression of Gpcr68 at various BOLT doses in activated T cells at acidic pH. (F) Anti-CD3 and anti-CD28 activated CD4 + T cells were treated with different doses of BOLT to determine the protein expression of GPCR68 using Western blot. (G-J) CCK8 assay was performed to analyze the effect of various pH on B16, MC38, 143B, and MG63 cell proliferation. (K-L) Effect of various doses of BOLT on the B16 and K7M2 cell growth to determine the IC-50 of BOLT. Error bars represent mean ± SEM. ∗∗ p < 0.01 and ∗ p < 0.05.

    Techniques Used: Zeta Potential Analyzer, Isolation, Quantitative RT-PCR, Expressing, Western Blot, CCK-8 Assay

    BOLT activates T cell PI3K-AKT-mTOR pathway to enhance T cell anti-tumor effect. (A) Flow cytometry plots compare IFN-γ and IL-2 expression at pH 7.8 and 6.0 along with various doses of BOLT in CD4 + T cells from CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (B, C) Bar graphs show IFN-γ and IL-2 expression in CD4 + T cells from CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (D) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 antibodies and incubated for 3 days with cell culture media of different pH levels. Western blot was performed to determine the phosphorylation of Akt and S6 under acidic conditions (pH 6.5) and alkaline pH (7.8). (E) Activated CD4 + T cells were treated with 0, 0.25, and 0.5 mg/mL doses of BOLT following CD4 + T cells activation at pH 7.8. Western blot analysis showing the phosphorylation of Akt and S6 were observed. (F) CD4 + T cells were activated and treated with BOLT at acidic pH. Western blot analysis was performed to determine the phosphorylation of Akt and S6. Two-way ANOVA was used for multiple comparisons. Experiments were conducted in triplicate. Data are mean ± SEM, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.
    Figure Legend Snippet: BOLT activates T cell PI3K-AKT-mTOR pathway to enhance T cell anti-tumor effect. (A) Flow cytometry plots compare IFN-γ and IL-2 expression at pH 7.8 and 6.0 along with various doses of BOLT in CD4 + T cells from CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (B, C) Bar graphs show IFN-γ and IL-2 expression in CD4 + T cells from CD4 Cre or GPCR68 fl/fl CD4 Cre mice. (D) Naïve CD4 + T cells were activated with anti-CD3 and anti-CD28 antibodies and incubated for 3 days with cell culture media of different pH levels. Western blot was performed to determine the phosphorylation of Akt and S6 under acidic conditions (pH 6.5) and alkaline pH (7.8). (E) Activated CD4 + T cells were treated with 0, 0.25, and 0.5 mg/mL doses of BOLT following CD4 + T cells activation at pH 7.8. Western blot analysis showing the phosphorylation of Akt and S6 were observed. (F) CD4 + T cells were activated and treated with BOLT at acidic pH. Western blot analysis was performed to determine the phosphorylation of Akt and S6. Two-way ANOVA was used for multiple comparisons. Experiments were conducted in triplicate. Data are mean ± SEM, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

    Techniques Used: Flow Cytometry, Expressing, Incubation, Cell Culture, Western Blot, Phospho-proteomics, Activation Assay

    Related Articles

    Isolation:

    Article Title: The mechanism of macrophage glycolysis regulating Th17 cell differentiation and promoting the progression of EAN mice.
    Article Snippet: This study investigates how macrophage glycolysis interacts with CD4+ T cells to drive experimental autoimmune neuritis (EAN) progression.. In EAN mice, expression of key glycolytic genes (LDHA, GLUT1, HK2, and PKM2) was gradually upregulated in splenic macrophages and sciatic nerve.. By the disease peak (day 18 p.i.), these genes reaching approximately 4-to-8-fold increase over controls, paralleling a continuous expansion of the splenic M1 macrophage population.

    Article Title: Regulation of Tph cell differentiation via farnesoid X receptor of dendritic cells in inflammatory bowel disease
    Article Snippet: .. After erythrocyte lysis, cell pellets were resuspended and naïve CD4 + T cells were isolated using the Naïve CD4 + T Cell Isolation Kit (130-104-453, Miltenyi, Germany) according to the manufacturer’s instructions. .. For the BMDC–T cell co-culture system, 3 × 10 5 naïve CD4 + T cells from WT or Fxr −/− mice were activated with plate-bound anti-CD3e (16–0031-85, eBioscience, 5ug/ml) for 24 h. They were then co-cultured with 3 × 10 4 BMDCs from WT or Fxr −/− mice with or without IL-12 (10 ng/mL, PeproTech, USA) in 96-well round-bottom plates for 72 h (37°C, 5% CO 2 ).

    Cell Isolation:

    Article Title: The mechanism of macrophage glycolysis regulating Th17 cell differentiation and promoting the progression of EAN mice.
    Article Snippet: This study investigates how macrophage glycolysis interacts with CD4+ T cells to drive experimental autoimmune neuritis (EAN) progression.. In EAN mice, expression of key glycolytic genes (LDHA, GLUT1, HK2, and PKM2) was gradually upregulated in splenic macrophages and sciatic nerve.. By the disease peak (day 18 p.i.), these genes reaching approximately 4-to-8-fold increase over controls, paralleling a continuous expansion of the splenic M1 macrophage population.

    Article Title: Prohibitin 2 is a key regulator of T cell proliferation, differentiation, and effector functions in vivo
    Article Snippet: .. CD4 + naïve T cells were purified using the naïve CD4 + T cell Isolation kit for mouse from Miltenyi Biotec (130-104-453). ..

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity
    Article Snippet: The mice were anesthetized intraperitoneally with tribromoethanol (# T48402 , Sigma). .. Naïve T cells were purified from lymph nodes as well as spleens of C57/BL6, CD4 Cre , GPCR68 fl/fl CD4 Cre (CKO) mice by using the mouse naïve CD4 + T Cell Isolation Kit (#130-104-453; Miltenyi Biotec) or naïve CD8 + T Cell Isolation Kit (#130-096-543; Miltenyi Biotec) for negative selection. .. Cells were stimulated with anti-CD3/CD28 antibodies (2 and 1 μg/well, respectively, #BE0001/BE0015, Bioxcell) and IL-2 (50U, #202-IL-050, R&D) in a 24-well plate in complete RPMI-1640 medium containing 10% FBS, 1% penicillin/streptomycin, 1% glutamine, 1% Sodium Pyruvate, 1% non-essential amino acids, and 0.1% β-mercaptoethanol for 3-5days.

    Article Title: Regulation of Tph cell differentiation via farnesoid X receptor of dendritic cells in inflammatory bowel disease
    Article Snippet: .. After erythrocyte lysis, cell pellets were resuspended and naïve CD4 + T cells were isolated using the Naïve CD4 + T Cell Isolation Kit (130-104-453, Miltenyi, Germany) according to the manufacturer’s instructions. .. For the BMDC–T cell co-culture system, 3 × 10 5 naïve CD4 + T cells from WT or Fxr −/− mice were activated with plate-bound anti-CD3e (16–0031-85, eBioscience, 5ug/ml) for 24 h. They were then co-cultured with 3 × 10 4 BMDCs from WT or Fxr −/− mice with or without IL-12 (10 ng/mL, PeproTech, USA) in 96-well round-bottom plates for 72 h (37°C, 5% CO 2 ).

    Selection:

    Article Title: The mechanism of macrophage glycolysis regulating Th17 cell differentiation and promoting the progression of EAN mice.
    Article Snippet: This study investigates how macrophage glycolysis interacts with CD4+ T cells to drive experimental autoimmune neuritis (EAN) progression.. In EAN mice, expression of key glycolytic genes (LDHA, GLUT1, HK2, and PKM2) was gradually upregulated in splenic macrophages and sciatic nerve.. By the disease peak (day 18 p.i.), these genes reaching approximately 4-to-8-fold increase over controls, paralleling a continuous expansion of the splenic M1 macrophage population.

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity
    Article Snippet: The mice were anesthetized intraperitoneally with tribromoethanol (# T48402 , Sigma). .. Naïve T cells were purified from lymph nodes as well as spleens of C57/BL6, CD4 Cre , GPCR68 fl/fl CD4 Cre (CKO) mice by using the mouse naïve CD4 + T Cell Isolation Kit (#130-104-453; Miltenyi Biotec) or naïve CD8 + T Cell Isolation Kit (#130-096-543; Miltenyi Biotec) for negative selection. .. Cells were stimulated with anti-CD3/CD28 antibodies (2 and 1 μg/well, respectively, #BE0001/BE0015, Bioxcell) and IL-2 (50U, #202-IL-050, R&D) in a 24-well plate in complete RPMI-1640 medium containing 10% FBS, 1% penicillin/streptomycin, 1% glutamine, 1% Sodium Pyruvate, 1% non-essential amino acids, and 0.1% β-mercaptoethanol for 3-5days.

    Purification:

    Article Title: Prohibitin 2 is a key regulator of T cell proliferation, differentiation, and effector functions in vivo
    Article Snippet: .. CD4 + naïve T cells were purified using the naïve CD4 + T cell Isolation kit for mouse from Miltenyi Biotec (130-104-453). ..

    Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity
    Article Snippet: The mice were anesthetized intraperitoneally with tribromoethanol (# T48402 , Sigma). .. Naïve T cells were purified from lymph nodes as well as spleens of C57/BL6, CD4 Cre , GPCR68 fl/fl CD4 Cre (CKO) mice by using the mouse naïve CD4 + T Cell Isolation Kit (#130-104-453; Miltenyi Biotec) or naïve CD8 + T Cell Isolation Kit (#130-096-543; Miltenyi Biotec) for negative selection. .. Cells were stimulated with anti-CD3/CD28 antibodies (2 and 1 μg/well, respectively, #BE0001/BE0015, Bioxcell) and IL-2 (50U, #202-IL-050, R&D) in a 24-well plate in complete RPMI-1640 medium containing 10% FBS, 1% penicillin/streptomycin, 1% glutamine, 1% Sodium Pyruvate, 1% non-essential amino acids, and 0.1% β-mercaptoethanol for 3-5days.

    Lysis:

    Article Title: Regulation of Tph cell differentiation via farnesoid X receptor of dendritic cells in inflammatory bowel disease
    Article Snippet: .. After erythrocyte lysis, cell pellets were resuspended and naïve CD4 + T cells were isolated using the Naïve CD4 + T Cell Isolation Kit (130-104-453, Miltenyi, Germany) according to the manufacturer’s instructions. .. For the BMDC–T cell co-culture system, 3 × 10 5 naïve CD4 + T cells from WT or Fxr −/− mice were activated with plate-bound anti-CD3e (16–0031-85, eBioscience, 5ug/ml) for 24 h. They were then co-cultured with 3 × 10 4 BMDCs from WT or Fxr −/− mice with or without IL-12 (10 ng/mL, PeproTech, USA) in 96-well round-bottom plates for 72 h (37°C, 5% CO 2 ).



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