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cd4 enrichment protocol  (STEMCELL Technologies Inc)

 
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    STEMCELL Technologies Inc cd4 enrichment protocol
    Cd4 Enrichment Protocol, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/cd4+enrichment+protocol/pmc05323111-171-9-19
    Average 90 stars, based on 1 article reviews
    cd4 enrichment protocol - by Bioz Stars, 2026-10
    90/100 stars

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    Article Title: Antigen specific immune response in Chlamydia muridarum genital infection is dependent on murine microRNAs-155 and -182
    Article Snippet: CD4 + T-cells were purified (purity > 85-90% by flow cytometry) using a CD4 enrichment protocol per manufacturer's recommendation (StemCell).



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    (A) Conventional T cells develop normally in the absence of p53. Upper panel, <t>CD4</t> and CD8 profile of total thymocytes. Lower panel, CD4 and CD8 profile of TCRβ high matured thymocytes. (B) Representative gating strategy for thymic NKT1 , NKT2 and NKT17 subsets. (C) p53 protein expression levels were analyzed by flow cytometry at steady state and after irradiation. For irradiation condition, cells were irradiated (1,000 rad) and incubated for 4 hours at 37°C. Data is pooled result of two independent experiments. (D) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data are mean ± SEM. Statistical differences between groups were analyzed with One-way ANOVA followed by multiple comparison. * * p < 0.005, * * * p < 0.0001.
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    (A) Conventional T cells develop normally in the absence of p53. Upper panel, <t>CD4</t> and CD8 profile of total thymocytes. Lower panel, CD4 and CD8 profile of TCRβ high matured thymocytes. (B) Representative gating strategy for thymic NKT1 , NKT2 and NKT17 subsets. (C) p53 protein expression levels were analyzed by flow cytometry at steady state and after irradiation. For irradiation condition, cells were irradiated (1,000 rad) and incubated for 4 hours at 37°C. Data is pooled result of two independent experiments. (D) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data are mean ± SEM. Statistical differences between groups were analyzed with One-way ANOVA followed by multiple comparison. * * p < 0.005, * * * p < 0.0001.
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    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from <t>2D2</t> mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.
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    (A) Conventional T cells develop normally in the absence of p53. Upper panel, CD4 and CD8 profile of total thymocytes. Lower panel, CD4 and CD8 profile of TCRβ high matured thymocytes. (B) Representative gating strategy for thymic NKT1 , NKT2 and NKT17 subsets. (C) p53 protein expression levels were analyzed by flow cytometry at steady state and after irradiation. For irradiation condition, cells were irradiated (1,000 rad) and incubated for 4 hours at 37°C. Data is pooled result of two independent experiments. (D) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data are mean ± SEM. Statistical differences between groups were analyzed with One-way ANOVA followed by multiple comparison. * * p < 0.005, * * * p < 0.0001.

    Journal: bioRxiv

    Article Title: Tumor suppressor p53 controls thymic NKT17 development

    doi: 10.1101/2024.08.21.608967

    Figure Lengend Snippet: (A) Conventional T cells develop normally in the absence of p53. Upper panel, CD4 and CD8 profile of total thymocytes. Lower panel, CD4 and CD8 profile of TCRβ high matured thymocytes. (B) Representative gating strategy for thymic NKT1 , NKT2 and NKT17 subsets. (C) p53 protein expression levels were analyzed by flow cytometry at steady state and after irradiation. For irradiation condition, cells were irradiated (1,000 rad) and incubated for 4 hours at 37°C. Data is pooled result of two independent experiments. (D) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data are mean ± SEM. Statistical differences between groups were analyzed with One-way ANOVA followed by multiple comparison. * * p < 0.005, * * * p < 0.0001.

    Article Snippet: Anti-CD4 (RM4-5)-BV510 (Need clone name or described it in Star Protocol), anti-B220 (RA3-6B2)-AF700, anti-CD44 (IM7)-FITC, anti-TCRβ (H57-597)-PE/Cy7, anti-RORγt (Q31-378)-BV786, anti-T-bet (O4-46)-BV421, anti-PLZF (R17-809)-AF647, anti-CCR7 (4B12)-BV786, anti-IL-17A (TC11-18H10)-FITC, anti-IFNγ (XMG1.2)-FITC, anti-IL-4 (11B11)-BV421 were purchased from BD Biosciences.

    Techniques: Expressing, Flow Cytometry, Irradiation, Incubation, Comparison

    (A) Ki67 expression in thymic iNKT subset was analyzed by flow cytometry. Data is pooled result of two independent experiments. WT, n=2; cKO, n=2. (B) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data is pooled result of three independent experiments. SP4; CD4 single positive thymocytes. WT, n=3; cKO, n=4. Data are mean ± SEM. Statistical differences were analyzed with student t-test. * p < 0.05.

    Journal: bioRxiv

    Article Title: Tumor suppressor p53 controls thymic NKT17 development

    doi: 10.1101/2024.08.21.608967

    Figure Lengend Snippet: (A) Ki67 expression in thymic iNKT subset was analyzed by flow cytometry. Data is pooled result of two independent experiments. WT, n=2; cKO, n=2. (B) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data is pooled result of three independent experiments. SP4; CD4 single positive thymocytes. WT, n=3; cKO, n=4. Data are mean ± SEM. Statistical differences were analyzed with student t-test. * p < 0.05.

    Article Snippet: Anti-CD4 (RM4-5)-BV510 (Need clone name or described it in Star Protocol), anti-B220 (RA3-6B2)-AF700, anti-CD44 (IM7)-FITC, anti-TCRβ (H57-597)-PE/Cy7, anti-RORγt (Q31-378)-BV786, anti-T-bet (O4-46)-BV421, anti-PLZF (R17-809)-AF647, anti-CCR7 (4B12)-BV786, anti-IL-17A (TC11-18H10)-FITC, anti-IFNγ (XMG1.2)-FITC, anti-IL-4 (11B11)-BV421 were purchased from BD Biosciences.

    Techniques: Expressing, Flow Cytometry

    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from 2D2 mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.

    Journal: Acta biomaterialia

    Article Title: Intrinsic immunogenicity of rapidly-degradable polymers evolves during degradation

    doi: 10.1016/j.actbio.2015.12.026

    Figure Lengend Snippet: Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from 2D2 mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.

    Article Snippet: After 48 hours in culture, CD4 + T cells were isolated from the spleens of 2D2 mice using the manufacturer’s recommended protocol (STEMCELL).

    Techniques: Immunopeptidomics, Isolation, Incubation, Flow Cytometry