cd48 antibody, anti-mouse  (Miltenyi Biotec)

Journal: iScience

Article Title: Lymphopoiesis is attenuated upon hepatocyte-specific deletion of the cytochrome c oxidase assembly factor Sco1

doi: 10.1016/j.isci.2025.112151

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Article Snippet: CD48-FITC (Clone: HM48-1) , ThermoFisher Scientific , Cat#: 11-0481-82; RRID: AB_465077.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Functional Assay, Western Blot, Recombinant, Protease Inhibitor, Diagnostic Assay, Staining, Software

Journal: iScience

Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and chimeric antigen receptor-T cells

doi: 10.1016/j.isci.2024.111669

Figure Lengend Snippet: 2B4 forms microclusters dependently on the binding to its ligand, CD48 (A) AND-TCR-Tg CD4 + T cells expressing m2B4-EGFP were plated onto an MCC 88–103 -prepulsed SLB containing I-E k -, mICAM-1-, and without or with mCD48-GPI. A representative of three independent experiments is shown. (B) Clustering and centripetal movement of 2B4 on the diagonal yellow lines in (A) are presented as horizontal elements in kymographs. (C) The cells in (A) were plated onto an MCC 88–103 -prepulsed or not prepulsed SLB without or with mCD48-GPI. Histograms show fold fluorescence intensities of TCRβ and 2B4 on the diagonal yellow lines in the differential interference contrast (DIC) images. A representative of five independent experiments is shown. (D) Percentages of T cells forming TCR or 2B4 microclusters at 2 min in (C) ( n = 50). (E) Scatterplot summarizing Pearson’s colocalization coefficient (PCC) values at 2 min in (C). PCC was calculated between 2B4/TCRβ by each randomly plotted profile on each cell (MCC 88–103 + CD48 − [left]; MCC 88–103 + CD48 + [middle]; MCC 88–103 − CD48 + [right]; 0.034 ± 0.09, 0.219 ± 0.16, 0.063 ± 0.11; n = 40, 101, 40; respectively). (F) OT-I-TCR-Tg effector CD8 + T cells non-transduced or transduced with m2B4 were co-cultured for 16 h with target mCD48-deleted EL-4 cells reconstituted by mCD48 (EL-4 ΔCd48 -CD48) not prepulsed or prepulsed by 1 μM OVA 257–264 . The percentage of specific lysis with and without 2B4 expression against OVA 257–264 -prepulsed target cells was statistically analyzed. A representative of three independent experiments is shown. (G) 2D12 cells not expressing or expressing m2B4-EGFP were plated onto an MCC 88–103 -prepulsed or not prepulsed SLB with mCD48-GPI. A representative of three independent experiments is shown. (H) The percentages of T cells forming pCD3ζ microclusters in (G) ( n = 120). (I) The scatterplot summarizes the PCC values between pCD3ζ/TCRβ in (G) (MCC + 2B4 − , MCC + 2B4 + , MCC − 2B4 + T cells; 0.398 ± 0.16, 0.402 ± 0.19, 0.058 ± 0.09; n = 28, 45, 45; respectively). (J) Cytokine secretion assay by ELISA. The T cells in (F) were co-cultured with mCD48-expressing EL-4 cells not prepulsed or prepulsed by 1 μM OVA 257–264 (left) or with splenocytes prepared from C57BL/6 mice and 1 nM OVA 257–264 (right) for 6 h. A representative of three independent experiments is shown. (K) 2D12 cells expressing m2B4 were stimulated by mCD48-deleted or mCD48-expressing EL-4 cells not prepulsed or prepulsed by 10 μM MCC 88–103 . A representative of three independent experiments is shown. (L) Each intensity ratio of pErk/Erk at 2 min for the case of MCC − 2B4 + in (K) is plotted on the graph. Bars, 5 μm. Data are presented as mean values ± SD. Statistical analysis was performed using a two-sided t -test or one-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. ns, not significant. See also Figure S1 .

Article Snippet: CD48 (HM48-1) , Bio X cell , BE0147; RRID: AB_10949470.

Techniques: Binding Assay, Expressing, Fluorescence, Transduction, Cell Culture, Lysis, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and chimeric antigen receptor-T cells

doi: 10.1016/j.isci.2024.111669

Figure Lengend Snippet: SHIP-1 colocalizes with 2B4 at TCR microcluster depending on the 2B4–CD48 binding (A) AND-TCR-Tg CD4 + T cells transduced with 2B4-EGFP and HaloTag-SHP-1, HaloTag-SHP-2 or SHIP-1-HaloTag were plated onto a not prepulsed SLB or MCC 88–103 -prepulsed SLB without or with CD48-GPI. A representative of four independent experiments is shown. (B) Percentages of T cells forming microclusters of each phosphatase at 2 min in (A) ( n = 50). (C) The PCC values of SHIP-1-expressing T cells plated on the MCC 88–103 -prepulsed SLB with CD48-GPI 2 min after contact in T cells of row 4 in (A). PCC was calculated between SHIP-1/TCRβ (0.138 ± 0.14, n = 105) or SHIP-1/2B4 (0.366 ± 0.22, n = 105). (D) 2D12 cells expressing 2B4-EGFP and HaloTag-SHP-1, HaloTag-SHP-2, or SHIP-1-HaloTag were conjugated with CD48-deleted or CD48-expressing EL-4 cells not prepulsed or prepulsed by MCC 88–103 . A representative of two independent experiments is shown. (E) 2D12 cells expressing SHIP-1-EGFP and 2B4-HaloTag were stimulated by mCD48-deleted or mCD48-expressing EL-4 cells not prepulsed or prepulsed by 10 μM MCC 88–103 . Lysates were immunoprecipitated for SHIP-1 by anti-GFP. A representative of three independent experiments is shown. (F) Each intensity ratio of 4G10/SHIP-1 (IP) at 2 min for the case of MCC + CD48 − in (E) is plotted on the graph. (G) 2D12 cells were transduced with SHIP-1-HaloTag and 2B4 (wild type)-EGFP or 2B4-4YF-EGFP and additionally introduced with SAP as indicated. These cells were plated onto an MCC 88–103 -prepulsed SLB without or with CD48-GPI. A representative of three independent experiments is shown. (H) Scatterplot summarizing the PCC values between 2B4/SHIP-1 in (G) (−0.016 ± 0.12, 0.005 ± 0.09, 0.233 ± 0.21, 0.027 ± 0.14, 0.064 ± 0.15; n = 22, 26, 80, 40, 28; respectively from the left of the graph).Bars, 5 μm. Data are presented as mean values ± SD. Statistical analysis was performed using two-sided t -test or one-way ANOVA. ∗ p < 0.05, ∗∗∗∗ p < 0.0001. See also Figure S2 .

Article Snippet: CD48 (HM48-1) , Bio X cell , BE0147; RRID: AB_10949470.

Techniques: Binding Assay, Transduction, Expressing, Immunoprecipitation

Journal: iScience

Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and chimeric antigen receptor-T cells

doi: 10.1016/j.isci.2024.111669

Figure Lengend Snippet: 2B4 recruits both SAP and Fyn at TCR-2B4 microclusters depending on binding to CD48 (A) AND-TCR-Tg T cells transduced with 2B4-EGFP and SAP-HaloTag were plated onto an MCC 88–103 -prepulsed SLB without or with CD48-GPI. A representative of four independent experiments is shown. (B) Percentages of T cells forming SAP microclusters at 2 min in (A) ( n = 70). (C) PCC was calculated between SAP/TCRβ (0.173 ± 0.14, n = 55) and SAP/2B4 (0.508 ± 0.23, n = 75) in T cells of the left bottom row in (A). (D) 2D12 cells transduced with SAP-HaloTag and wild type (WT) 2B4-EGFP or 2B4-4YF-EGFP were plated onto an MCC 88–103 -prepulsed SLB with CD48-GPI. A representative of two independent experiments is shown. (E) Percentages of T cells forming SAP microclusters in (D) ( n = 30). (F) 2D12 cells expressing 2B4-EGFP and Fyn-HaloTag were not transduced or transduced with SAP and were stimulated by mCD48-deleted or mCD48-expressing EL-4 cells prepulsed by MCC 88–103 . Lysates were immunoprecipitated for 2B4 by anti-GFP. A representative of three independent experiments is shown. (G) Intensities of SAP or Fyn in (F) are shown. (H) The T cells in (F) were plated onto an MCC 88–103 -prepulsed SLB without or with CD48-GPI. A representative of three independent experiments is shown. (I) Percentages of T cells forming Fyn microclusters in (H) ( n = 70). (J) PCC was calculated between Fyn/TCRβ (0.360 ± 0.16, n = 72) and Fyn/2B4 (0.622 ± 0.14, n = 72) in T cells of row 3 in (H). (K) IL-2 secretion assay by ELISA. OT91 cells non-transduced or transduced with 2B4 alone or both 2B4 and SAP were co-cultured with mCD48-deleted or mCD48-expressing EL-4 cells prepulsed with OVA 257–264 . A representative of three independent experiments is shown. (L) 2D12 cells non-transduced or transduced with WT 2B4, 2B4-4YF, or both WT 2B4 and SAP were stimulated by mCD48-expressing EL-4 cells prepulsed by MCC 88–103 . A representative of three independent experiments is shown. (M) Each intensity ratio of pErk/Erk at 2 min or pVav1/Vav1 at 0.5 min for the case of 2B4 − SAP − in (L) is plotted on the graph. (N) The schematic model of 2B4 signaling pathways without CD48 on target cells (left), with CD48 on target cells and little SAP in T cells (middle), and with CD48 on target cells and sufficient SAP in T cells (right). Bars, 5 μm. Data are presented as mean values ± SD. Statistical analysis was performed by two-sided t -test or one-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. ns, not significant. See also Figure S3 .

Article Snippet: CD48 (HM48-1) , Bio X cell , BE0147; RRID: AB_10949470.

Techniques: Binding Assay, Transduction, Expressing, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Cell Culture, Protein-Protein interactions

Journal: iScience

Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and chimeric antigen receptor-T cells

doi: 10.1016/j.isci.2024.111669

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Article Snippet: CD48 (HM48-1) , Bio X cell , BE0147; RRID: AB_10949470.

Techniques: Control, Purification, Virus, Recombinant, Isolation, Enzyme-linked Immunosorbent Assay, Labeling, Retroviral, Plasmid Preparation, Luciferase, Software

Journal: Chinese Medicine

Article Title: Siwu decoction mitigates radiation-induced immune senescence by attenuating hematopoietic damage

doi: 10.1186/s13020-024-01036-3

Figure Lengend Snippet: Antibodies used in flow cytometry

Article Snippet: CD48 , HM48-1 , APC-EF780 , eBioscience , 47–0481–82.

Techniques: