ceacam1 protein human recombinant  (Sino Biological)


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    Name:
    Human CEACAM1 CD66a Protein His Tag
    Description:
    A DNA sequence encoding the extracellular domain of human CEACAM1 NP 001020083 1 Met 1 Gly 428 was expressed with a C terminal polyhistidine tag
    Catalog Number:
    10822-H08H
    Price:
    None
    Category:
    recombinant protein
    Host:
    HEK293 Cells
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    Structured Review

    Sino Biological ceacam1 protein human recombinant
    A DNA sequence encoding the extracellular domain of human CEACAM1 NP 001020083 1 Met 1 Gly 428 was expressed with a C terminal polyhistidine tag
    https://www.bioz.com/result/ceacam1 protein human recombinant/product/Sino Biological
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ceacam1 protein human recombinant - by Bioz Stars, 2021-07
    91/100 stars

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    Article Title: Thioloxidoreductase HP0231 of Helicobacter pylori impacts HopQ-dependent CagA translocation.
    Article Snippet: Bacterial pull downAnalysis of the HopQ and its mutated variants interactions with human His-tagged CEACAM1 (10822-H08H, Sino Biological) by pulldown experiment was performed as described (Javaheri et al., 2016).

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  • 93
    Sino Biological human ceacam1
    Ligand-blocking properties of an anti-human TIM-3 antibody. A, hTIM-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to ELISA plates coated with galectin-9. B, hTim-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to dexamethasone treated thymocytes. C, Untransduced or human <t>CEACAM1-transduced</t> Jurkat T cells were stained with hTim-3-Ig that was pre-incubated with no antibody, F38.2E2 or matched isotype control antibody. Data are representative of 6 independent experiments.
    Human Ceacam1, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human ceacam1/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human ceacam1 - by Bioz Stars, 2021-07
    93/100 stars
      Buy from Supplier

    90
    Sino Biological mouse ceacam1
    Ligand-blocking properties of an anti-human TIM-3 antibody. A, hTIM-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to ELISA plates coated with galectin-9. B, hTim-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to dexamethasone treated thymocytes. C, Untransduced or human <t>CEACAM1-transduced</t> Jurkat T cells were stained with hTim-3-Ig that was pre-incubated with no antibody, F38.2E2 or matched isotype control antibody. Data are representative of 6 independent experiments.
    Mouse Ceacam1, supplied by Sino Biological, 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/result/mouse ceacam1/product/Sino Biological
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse ceacam1 - by Bioz Stars, 2021-07
    90/100 stars
      Buy from Supplier

    N/A
    Produced in rabbits immunized with purified recombinant Human CEACAM1 extracellular domain rhCEACAM1 Catalog 10822 H08H NP 001020083 1 Met 1 Gly 428 Total IgG was purified by Protein A affinity
      Buy from Supplier

    N/A
    This antibody was obtained from a rabbit immunized with purified recombinant Human CEACAM‑1 CD66a rh CEACAM‑1 CD66a Catalog 10822 H08H NP 001020083 1 Met1 Gly428
      Buy from Supplier

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    Ligand-blocking properties of an anti-human TIM-3 antibody. A, hTIM-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to ELISA plates coated with galectin-9. B, hTim-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to dexamethasone treated thymocytes. C, Untransduced or human CEACAM1-transduced Jurkat T cells were stained with hTim-3-Ig that was pre-incubated with no antibody, F38.2E2 or matched isotype control antibody. Data are representative of 6 independent experiments.

    Journal: Oncoimmunology

    Article Title: Blockade of Tim-3 binding to phosphatidylserine and CEACAM1 is a shared feature of anti-Tim-3 antibodies that have functional efficacy

    doi: 10.1080/2162402X.2017.1385690

    Figure Lengend Snippet: Ligand-blocking properties of an anti-human TIM-3 antibody. A, hTIM-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to ELISA plates coated with galectin-9. B, hTim-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to dexamethasone treated thymocytes. C, Untransduced or human CEACAM1-transduced Jurkat T cells were stained with hTim-3-Ig that was pre-incubated with no antibody, F38.2E2 or matched isotype control antibody. Data are representative of 6 independent experiments.

    Article Snippet: For detection of the interaction between human CEACAM1 and human Tim-3, 10 μg/ml recombinant human Tim-3-Ig harboring a human IgG1 tail (Sino Biological) was incubated with 10 μg/ml of either anti-human Tim-3 (F38.2E2) or isotype control.

    Techniques: Blocking Assay, Incubation, Enzyme-linked Immunosorbent Assay, Staining

    Effect of anti-murine Tim-3 antibodies on binding to CEACAM1. A, Expression of murine CEACAM1 on transduced Jurkat T cells. B, Untransduced or murine CEACAM1-transduced Jurkat T cells were stained with mTim-3-Ig that was pre-incubated with no antibody, RMT3-23, 2C12, 5D12 or matched isotype control antibody. Data are representative of 3 independent experiments.

    Journal: Oncoimmunology

    Article Title: Blockade of Tim-3 binding to phosphatidylserine and CEACAM1 is a shared feature of anti-Tim-3 antibodies that have functional efficacy

    doi: 10.1080/2162402X.2017.1385690

    Figure Lengend Snippet: Effect of anti-murine Tim-3 antibodies on binding to CEACAM1. A, Expression of murine CEACAM1 on transduced Jurkat T cells. B, Untransduced or murine CEACAM1-transduced Jurkat T cells were stained with mTim-3-Ig that was pre-incubated with no antibody, RMT3-23, 2C12, 5D12 or matched isotype control antibody. Data are representative of 3 independent experiments.

    Article Snippet: For detection of the interaction between human CEACAM1 and human Tim-3, 10 μg/ml recombinant human Tim-3-Ig harboring a human IgG1 tail (Sino Biological) was incubated with 10 μg/ml of either anti-human Tim-3 (F38.2E2) or isotype control.

    Techniques: Binding Assay, Expressing, Staining, Incubation

    TIM‐3 and CEACAM1 do not interact in cis . (A) HEK293T cells were cotransfected with indicated molecules. Flow cytometry analysis was performed 2 days after transfection. Open histograms: staining with an isotype control; filled histograms: staining of indicated molecules. Bar diagrams show results of three independent experiments. (B) Analysis of in cis association of indicated interaction partners via FRET. mRuby3 (561 nm excitation; PE filter) and mNeonGreen (488 nm excitation; FITC filter) fusion constructs were cotransfected in HEK293T cells and analyzed by flow cytometry. For FRET detection, 488 nm laser light was used for fluorophore excitation, and mRuby3 emission (600 nm longpass filter) was measured. (C) Fluorescence images of HEK293T cells coexpressing the indicated molecules. FRET yield was determined by donor recovery after acceptor photobleaching and calculated pixelwise. Representative images are shown. Scale bar = 5 μm. (D) Quantification of average FRET yield per cell ( n = 46; 35; 57). For statistical evaluation, one‐way ANOVA followed by Tukey's test was performed (ns, p > 0.05; **** p ≤ 0.0001). (A and D) Standard error of the mean is shown.

    Journal: European Journal of Immunology

    Article Title: TIM‐3 and CEACAM1 do not interact in cis and in trans

    doi: 10.1002/eji.201948400

    Figure Lengend Snippet: TIM‐3 and CEACAM1 do not interact in cis . (A) HEK293T cells were cotransfected with indicated molecules. Flow cytometry analysis was performed 2 days after transfection. Open histograms: staining with an isotype control; filled histograms: staining of indicated molecules. Bar diagrams show results of three independent experiments. (B) Analysis of in cis association of indicated interaction partners via FRET. mRuby3 (561 nm excitation; PE filter) and mNeonGreen (488 nm excitation; FITC filter) fusion constructs were cotransfected in HEK293T cells and analyzed by flow cytometry. For FRET detection, 488 nm laser light was used for fluorophore excitation, and mRuby3 emission (600 nm longpass filter) was measured. (C) Fluorescence images of HEK293T cells coexpressing the indicated molecules. FRET yield was determined by donor recovery after acceptor photobleaching and calculated pixelwise. Representative images are shown. Scale bar = 5 μm. (D) Quantification of average FRET yield per cell ( n = 46; 35; 57). For statistical evaluation, one‐way ANOVA followed by Tukey's test was performed (ns, p > 0.05; **** p ≤ 0.0001). (A and D) Standard error of the mean is shown.

    Article Snippet: TIM‐3 and CEACAM1 Ig fusion proteins (TIM‐3‐Ig, CEACAM1‐Ig) were purchased from SinoBiological (SB; People's Republic of China).

    Techniques: Flow Cytometry, Transfection, Staining, Construct, Fluorescence

    TIM‐3 does not modulate CEACAM1 function. (A) Flow cytometry analysis of TPR reporter cells coexpressing CEACAM1‐4L and TIM‐3 (gray histograms). Reactivity of antibodies to CEACAM1 and TIM‐3 to control TPR is shown as open histograms. (B) CEACAM1‐4L reporter cells coexpressing TIM‐3 were stimulated with control TCS or TCS‐expressing CEACAM1. Results are from seven independent experiments performed in triplicates. (C) Expression of membrane‐bound anti‐CD3 Ab fragment and TIM‐3 on TCS‐TIM‐3 (gray histograms). Reactivity of the used antibodies to control cells is shown as open histograms. (D) The indicated TPRs were stimulated with control TCS or TCS‐expressing TIM‐3. Results are from five independent experiments performed in triplicates. (B and D) eGFP, eCFP, and mCherry expression was measured via flow cytometry. Reporter activation is shown as fold induction (gMFI of TCS‐stimulated cells/gMFI of unstimulated cells). Note that some data points overlap. For statistical evaluation, paired t ‐tests were performed (**** p ≤ 0.0001; ** p ≤ 0.01; ns, p > 0.05).

    Journal: European Journal of Immunology

    Article Title: TIM‐3 and CEACAM1 do not interact in cis and in trans

    doi: 10.1002/eji.201948400

    Figure Lengend Snippet: TIM‐3 does not modulate CEACAM1 function. (A) Flow cytometry analysis of TPR reporter cells coexpressing CEACAM1‐4L and TIM‐3 (gray histograms). Reactivity of antibodies to CEACAM1 and TIM‐3 to control TPR is shown as open histograms. (B) CEACAM1‐4L reporter cells coexpressing TIM‐3 were stimulated with control TCS or TCS‐expressing CEACAM1. Results are from seven independent experiments performed in triplicates. (C) Expression of membrane‐bound anti‐CD3 Ab fragment and TIM‐3 on TCS‐TIM‐3 (gray histograms). Reactivity of the used antibodies to control cells is shown as open histograms. (D) The indicated TPRs were stimulated with control TCS or TCS‐expressing TIM‐3. Results are from five independent experiments performed in triplicates. (B and D) eGFP, eCFP, and mCherry expression was measured via flow cytometry. Reporter activation is shown as fold induction (gMFI of TCS‐stimulated cells/gMFI of unstimulated cells). Note that some data points overlap. For statistical evaluation, paired t ‐tests were performed (**** p ≤ 0.0001; ** p ≤ 0.01; ns, p > 0.05).

    Article Snippet: TIM‐3 and CEACAM1 Ig fusion proteins (TIM‐3‐Ig, CEACAM1‐Ig) were purchased from SinoBiological (SB; People's Republic of China).

    Techniques: Flow Cytometry, Expressing, Activation Assay

    TIM‐3 and CEACAM1 do not interact in trans . (A) Binding of indicated Ig fusion proteins to immobilized recombinant galectin‐9 (Gal‐9) and CEACAM1 proteins was analyzed by ELISA. Results are representative for four independently performed experiments. Information on the used proteins is provided in the material and method section. (B) Jurkat cells transduced to express ICOS, TIM‐3, or CEACAM1 were probed with the respective antibodies and analyzed by flow cytometry (gray histograms). Open histograms show the reactivity of control Jurkat cells. Bottom: results of a representative binding experiment of indicated Ig fusion proteins used at a final concentration of 10 μg/mL to control Jurkat cells or Jurkat cells expressing ICOS, TIM‐3, or CEACAM1. Binding was detected via flow cytometry by an APC‐conjugated goat‐anti‐human IgG (Fc‐specific) Ab. (C) Binding of indicated Ig fusion proteins (final concentrations: 31.6, 10, 3.16, 1, and 0.316 μg/mL) to control Jurkat cells and Jurkat cells expressing ICOS, TIM‐3, or CEACAM1. Binding was detected as described in (B). Results are shown from three different experiments performed in duplicates. Binding signals (gMFI) were normalized to background binding (gMFI values obtained with secondary reagent only). (D) Binding of CEACAM1‐Ig (final concentration: 10 μg/mL) to control Jurkat cells and CEACAM1‐expressing Jurkat cells in absence or presence of a blocking CEACAM1 mAb. Results are shown from four different experiments performed in duplicates. (E) Cells coexpressing eCFP or eGFP and the indicated cell surface molecules were preincubated for 1 h and conjugate formation between cells expressing different fluorescent proteins was assessed via flow cytometry. (F) Results of three independently performed cell–cell binding assays are summarized. (A, C, D, and F) Standard deviation is shown.

    Journal: European Journal of Immunology

    Article Title: TIM‐3 and CEACAM1 do not interact in cis and in trans

    doi: 10.1002/eji.201948400

    Figure Lengend Snippet: TIM‐3 and CEACAM1 do not interact in trans . (A) Binding of indicated Ig fusion proteins to immobilized recombinant galectin‐9 (Gal‐9) and CEACAM1 proteins was analyzed by ELISA. Results are representative for four independently performed experiments. Information on the used proteins is provided in the material and method section. (B) Jurkat cells transduced to express ICOS, TIM‐3, or CEACAM1 were probed with the respective antibodies and analyzed by flow cytometry (gray histograms). Open histograms show the reactivity of control Jurkat cells. Bottom: results of a representative binding experiment of indicated Ig fusion proteins used at a final concentration of 10 μg/mL to control Jurkat cells or Jurkat cells expressing ICOS, TIM‐3, or CEACAM1. Binding was detected via flow cytometry by an APC‐conjugated goat‐anti‐human IgG (Fc‐specific) Ab. (C) Binding of indicated Ig fusion proteins (final concentrations: 31.6, 10, 3.16, 1, and 0.316 μg/mL) to control Jurkat cells and Jurkat cells expressing ICOS, TIM‐3, or CEACAM1. Binding was detected as described in (B). Results are shown from three different experiments performed in duplicates. Binding signals (gMFI) were normalized to background binding (gMFI values obtained with secondary reagent only). (D) Binding of CEACAM1‐Ig (final concentration: 10 μg/mL) to control Jurkat cells and CEACAM1‐expressing Jurkat cells in absence or presence of a blocking CEACAM1 mAb. Results are shown from four different experiments performed in duplicates. (E) Cells coexpressing eCFP or eGFP and the indicated cell surface molecules were preincubated for 1 h and conjugate formation between cells expressing different fluorescent proteins was assessed via flow cytometry. (F) Results of three independently performed cell–cell binding assays are summarized. (A, C, D, and F) Standard deviation is shown.

    Article Snippet: TIM‐3 and CEACAM1 Ig fusion proteins (TIM‐3‐Ig, CEACAM1‐Ig) were purchased from SinoBiological (SB; People's Republic of China).

    Techniques: Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Concentration Assay, Expressing, Blocking Assay, Standard Deviation

    Evaluation of CEACAM1 function in a reporter cell system. (A) Schematic of CEACAM1‐4L and CEACAM1‐4S proteins. (B) Flow cytometry analysis of triple parameter reporter cells (TPR) and T cell stimulator cells (TCS). Open histograms: control cells; filled histograms: expression of indicated molecules on TPR and TCS. (C) Gating strategy and one representative stimulation experiment of control TPR and TPR‐expressing CEACAM1‐4S and CEACAM1‐4L with control TCS or TCS‐expressing CEACAM1 is shown. eGFP, eCFP, and mCherry expression was measured via flow cytometry. The histograms of unstimulated cells are also depicted. The geometric MFI (gMFI) value is shown for each histogram. (D) The indicated TPRs were stimulated with control TCS or TCS‐expressing CEACAM1. Reporter activation is shown as fold induction (gMFI of TCS‐stimulated cells/gMFI of unstimulated cells). Results are from ten independent experiments performed in triplicates. Note that some data points overlap. For statistical evaluation, paired t ‐tests were performed (**** p ≤ 0.0001; ** p ≤ 0.01; * p ≤ 0.05; ns, p > 0.05).

    Journal: European Journal of Immunology

    Article Title: TIM‐3 and CEACAM1 do not interact in cis and in trans

    doi: 10.1002/eji.201948400

    Figure Lengend Snippet: Evaluation of CEACAM1 function in a reporter cell system. (A) Schematic of CEACAM1‐4L and CEACAM1‐4S proteins. (B) Flow cytometry analysis of triple parameter reporter cells (TPR) and T cell stimulator cells (TCS). Open histograms: control cells; filled histograms: expression of indicated molecules on TPR and TCS. (C) Gating strategy and one representative stimulation experiment of control TPR and TPR‐expressing CEACAM1‐4S and CEACAM1‐4L with control TCS or TCS‐expressing CEACAM1 is shown. eGFP, eCFP, and mCherry expression was measured via flow cytometry. The histograms of unstimulated cells are also depicted. The geometric MFI (gMFI) value is shown for each histogram. (D) The indicated TPRs were stimulated with control TCS or TCS‐expressing CEACAM1. Reporter activation is shown as fold induction (gMFI of TCS‐stimulated cells/gMFI of unstimulated cells). Results are from ten independent experiments performed in triplicates. Note that some data points overlap. For statistical evaluation, paired t ‐tests were performed (**** p ≤ 0.0001; ** p ≤ 0.01; * p ≤ 0.05; ns, p > 0.05).

    Article Snippet: TIM‐3 and CEACAM1 Ig fusion proteins (TIM‐3‐Ig, CEACAM1‐Ig) were purchased from SinoBiological (SB; People's Republic of China).

    Techniques: Flow Cytometry, Expressing, Activation Assay

    CEACAM1 does not mediate TIM‐3 signaling. (A) Schematic of WT and mutated TIM‐3 molecules (left) and flow cytometric analysis of TPR reporter cells expressing the indicated TIM‐3 molecules (gray histograms) and control TPR (open histogram). (B) The indicated TPR were stimulated with control TCS or TCS‐expressing CEACAM1. eGFP, eCFP, and mCherry expression was measured via flow cytometry. Results are shown for five independent experiments performed in triplicates. Reporter activation is shown as fold induction (gMFI of TCS‐stimulated cells/gMFI of unstimulated cells). For statistical evaluation, unpaired t ‐tests were performed (ns, p > 0.05).

    Journal: European Journal of Immunology

    Article Title: TIM‐3 and CEACAM1 do not interact in cis and in trans

    doi: 10.1002/eji.201948400

    Figure Lengend Snippet: CEACAM1 does not mediate TIM‐3 signaling. (A) Schematic of WT and mutated TIM‐3 molecules (left) and flow cytometric analysis of TPR reporter cells expressing the indicated TIM‐3 molecules (gray histograms) and control TPR (open histogram). (B) The indicated TPR were stimulated with control TCS or TCS‐expressing CEACAM1. eGFP, eCFP, and mCherry expression was measured via flow cytometry. Results are shown for five independent experiments performed in triplicates. Reporter activation is shown as fold induction (gMFI of TCS‐stimulated cells/gMFI of unstimulated cells). For statistical evaluation, unpaired t ‐tests were performed (ns, p > 0.05).

    Article Snippet: TIM‐3 and CEACAM1 Ig fusion proteins (TIM‐3‐Ig, CEACAM1‐Ig) were purchased from SinoBiological (SB; People's Republic of China).

    Techniques: Expressing, Flow Cytometry, Activation Assay

    Cytoplasmic sequences of TIM‐3 and CEACAM1 induce inhibitory signals. (A) Schematic of mICOS chimera; (B) Left: surface expression of mICOS::Δcyt, mICOS::PD‐1, mICOS::TIM‐3, mICOS::TIM‐3_mut, mICOS::CEACAM1‐L, and mICOS::CEACAM1‐S on TPR cells (dark gray histograms). Reactivity of mICOS Ab with control TPR is shown as an open histogram. Right: Flow cytometry analysis of TCS‐mICOS‐L. Open histograms: control cells; filled histograms: expression of indicated molecules. (C) mICOS::Δcyt, mICOS::PD‐1, mICOS::TIM‐3, mICOS::TIM‐3_mut, mICOS::CEACAM1‐L, and mICOS::CEACAM1‐S reporter cells were stimulated with control TCS or TCS‐expressing mICOS‐L. For statistical evaluation, one‐way ANOVA followed by Dunnett's test was performed (**** p ≤ 0.0001; ns, p > 0.05). eGFP, eCFP, and mCherry expression was measured via flow cytometry. Results are shown for four independent experiments performed in triplicates. Reporter activation is shown as fold induction (gMFI of mICOS‐L‐stimulated cells/gMFI of control‐stimulated cells). It should be noted that some data pointes overlap. Control stimulation is shown as a dotted line.

    Journal: European Journal of Immunology

    Article Title: TIM‐3 and CEACAM1 do not interact in cis and in trans

    doi: 10.1002/eji.201948400

    Figure Lengend Snippet: Cytoplasmic sequences of TIM‐3 and CEACAM1 induce inhibitory signals. (A) Schematic of mICOS chimera; (B) Left: surface expression of mICOS::Δcyt, mICOS::PD‐1, mICOS::TIM‐3, mICOS::TIM‐3_mut, mICOS::CEACAM1‐L, and mICOS::CEACAM1‐S on TPR cells (dark gray histograms). Reactivity of mICOS Ab with control TPR is shown as an open histogram. Right: Flow cytometry analysis of TCS‐mICOS‐L. Open histograms: control cells; filled histograms: expression of indicated molecules. (C) mICOS::Δcyt, mICOS::PD‐1, mICOS::TIM‐3, mICOS::TIM‐3_mut, mICOS::CEACAM1‐L, and mICOS::CEACAM1‐S reporter cells were stimulated with control TCS or TCS‐expressing mICOS‐L. For statistical evaluation, one‐way ANOVA followed by Dunnett's test was performed (**** p ≤ 0.0001; ns, p > 0.05). eGFP, eCFP, and mCherry expression was measured via flow cytometry. Results are shown for four independent experiments performed in triplicates. Reporter activation is shown as fold induction (gMFI of mICOS‐L‐stimulated cells/gMFI of control‐stimulated cells). It should be noted that some data pointes overlap. Control stimulation is shown as a dotted line.

    Article Snippet: TIM‐3 and CEACAM1 Ig fusion proteins (TIM‐3‐Ig, CEACAM1‐Ig) were purchased from SinoBiological (SB; People's Republic of China).

    Techniques: Expressing, Flow Cytometry, Activation Assay

    Expression analysis of TIM‐3 and CEACAM1. Flow cytometry analysis of TIM‐3 expression on freshly isolated PBMCs derived from healthy donors. CD19 + cells (A), CD56 + cells (B), and CD14 + cells (C) were analyzed for TIM‐3 expression. (D) Immature (iDC) and mature DCs were stained with TIM‐3 mAb (black histograms) or isotype control (open histograms). (A–D) Left: one representative donor; right: each dot represents one donor. Median is shown. (E and F) CD4 + and CD8 + T cells in freshly isolated PBMCs and PBMCs stimulated in vitro with staphylococcal enterotoxin E (SEE) (E) or immobilized aCD3/aCD28 mAb (F) for 3, 6, and 10 days were analyzed for TIM‐3, CEACAM1, and CD25 expression, respectively. In the stimulated samples, the gate was set on proliferated (CFSE low ) cells. Left panels: dot plots from one representative donor are shown. For better visibility larger dots were used in dot plots depicting TIM‐3 and CEACAM1 expression in CD8 + T cells upon stimulation with SEE. Middle‐right panels: histogram overlay shows gMFI of CD25 expression on day 10 of the indicated populations. Right panels: summarized data of CD25 expression in the TIM3 + versus TIM3 + /CD66a + subset upon SEE (17 donors/8 experiments with 1–3 donors each) or aCD3/aCD28‐stimulation (11 donors/6 experiments with 1–3 donors each). (G) Bar diagrams from five representative donors (from three experiments with one or two donors) show percentages of CD66a + ‐expressing cells within the TIM3 + population. For statistical evaluation, paired t ‐tests (A–C; E and F) and one‐way ANOVA followed by Tukey's multiple comparison test (D) were performed (*** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05; ns, p > 0.05).

    Journal: European Journal of Immunology

    Article Title: TIM‐3 and CEACAM1 do not interact in cis and in trans

    doi: 10.1002/eji.201948400

    Figure Lengend Snippet: Expression analysis of TIM‐3 and CEACAM1. Flow cytometry analysis of TIM‐3 expression on freshly isolated PBMCs derived from healthy donors. CD19 + cells (A), CD56 + cells (B), and CD14 + cells (C) were analyzed for TIM‐3 expression. (D) Immature (iDC) and mature DCs were stained with TIM‐3 mAb (black histograms) or isotype control (open histograms). (A–D) Left: one representative donor; right: each dot represents one donor. Median is shown. (E and F) CD4 + and CD8 + T cells in freshly isolated PBMCs and PBMCs stimulated in vitro with staphylococcal enterotoxin E (SEE) (E) or immobilized aCD3/aCD28 mAb (F) for 3, 6, and 10 days were analyzed for TIM‐3, CEACAM1, and CD25 expression, respectively. In the stimulated samples, the gate was set on proliferated (CFSE low ) cells. Left panels: dot plots from one representative donor are shown. For better visibility larger dots were used in dot plots depicting TIM‐3 and CEACAM1 expression in CD8 + T cells upon stimulation with SEE. Middle‐right panels: histogram overlay shows gMFI of CD25 expression on day 10 of the indicated populations. Right panels: summarized data of CD25 expression in the TIM3 + versus TIM3 + /CD66a + subset upon SEE (17 donors/8 experiments with 1–3 donors each) or aCD3/aCD28‐stimulation (11 donors/6 experiments with 1–3 donors each). (G) Bar diagrams from five representative donors (from three experiments with one or two donors) show percentages of CD66a + ‐expressing cells within the TIM3 + population. For statistical evaluation, paired t ‐tests (A–C; E and F) and one‐way ANOVA followed by Tukey's multiple comparison test (D) were performed (*** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05; ns, p > 0.05).

    Article Snippet: TIM‐3 and CEACAM1 Ig fusion proteins (TIM‐3‐Ig, CEACAM1‐Ig) were purchased from SinoBiological (SB; People's Republic of China).

    Techniques: Expressing, Flow Cytometry, Isolation, Derivative Assay, Staining, In Vitro

    Ligand-blocking properties of an anti-human TIM-3 antibody. A, hTIM-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to ELISA plates coated with galectin-9. B, hTim-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to dexamethasone treated thymocytes. C, Untransduced or human CEACAM1-transduced Jurkat T cells were stained with hTim-3-Ig that was pre-incubated with no antibody, F38.2E2 or matched isotype control antibody. Data are representative of 6 independent experiments.

    Journal: Oncoimmunology

    Article Title: Blockade of Tim-3 binding to phosphatidylserine and CEACAM1 is a shared feature of anti-Tim-3 antibodies that have functional efficacy

    doi: 10.1080/2162402X.2017.1385690

    Figure Lengend Snippet: Ligand-blocking properties of an anti-human TIM-3 antibody. A, hTIM-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to ELISA plates coated with galectin-9. B, hTim-3-Ig was incubated with F38.2E2 or matched isotype control antibody prior to addition to dexamethasone treated thymocytes. C, Untransduced or human CEACAM1-transduced Jurkat T cells were stained with hTim-3-Ig that was pre-incubated with no antibody, F38.2E2 or matched isotype control antibody. Data are representative of 6 independent experiments.

    Article Snippet: To detect interaction between mouse CEACAM1 and Tim-3, 10 μg/ml recombinant mouse Tim-3-Ig fusion protein harboring a human IgG1 tail (Chimerigen) was incubated with 10 μg/ml of either anti-murine Tim-3 antibody or isotype control antibody for 30 minutes at room temperature prior to addition to the transfected cells.

    Techniques: Blocking Assay, Incubation, Enzyme-linked Immunosorbent Assay, Staining

    Effect of anti-murine Tim-3 antibodies on binding to CEACAM1. A, Expression of murine CEACAM1 on transduced Jurkat T cells. B, Untransduced or murine CEACAM1-transduced Jurkat T cells were stained with mTim-3-Ig that was pre-incubated with no antibody, RMT3-23, 2C12, 5D12 or matched isotype control antibody. Data are representative of 3 independent experiments.

    Journal: Oncoimmunology

    Article Title: Blockade of Tim-3 binding to phosphatidylserine and CEACAM1 is a shared feature of anti-Tim-3 antibodies that have functional efficacy

    doi: 10.1080/2162402X.2017.1385690

    Figure Lengend Snippet: Effect of anti-murine Tim-3 antibodies on binding to CEACAM1. A, Expression of murine CEACAM1 on transduced Jurkat T cells. B, Untransduced or murine CEACAM1-transduced Jurkat T cells were stained with mTim-3-Ig that was pre-incubated with no antibody, RMT3-23, 2C12, 5D12 or matched isotype control antibody. Data are representative of 3 independent experiments.

    Article Snippet: To detect interaction between mouse CEACAM1 and Tim-3, 10 μg/ml recombinant mouse Tim-3-Ig fusion protein harboring a human IgG1 tail (Chimerigen) was incubated with 10 μg/ml of either anti-murine Tim-3 antibody or isotype control antibody for 30 minutes at room temperature prior to addition to the transfected cells.

    Techniques: Binding Assay, Expressing, Staining, Incubation