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R&D Systems ic
Ic, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cd54/Recombinant+Mouse+ICAM-1%2FCD54+Fc+Chimera+Protein%2C+CF/pm41964950-520-32-40
Average 94 stars, based on 77 article reviews

ic - by Bioz Stars, 2026-09

94/100 stars

Images

Immunostaining:

Article Title: Methods and compositions for treatment of atherosclerosis
Article Snippet: .. Quantitative immunostaining was performed using primary antibodies against Mac-3 (1:200; BD PharmingenTM), α-smooth muscle actin (1:200; Abcam), CD106 (1:40; BD PharmingenTM) or CD54 (1:40; R&D systems), and fluorescently-labeled secondary antibodies goat anti-rat Alexa Fluor 488 (ThermoFisher Scientific, A11006), donkey anti-goat Alexa Fluor 594 (ThermoFisher Scientific, A11058), or goat anti-rabbit Alexa Fluor 594 (ThermoFisher Scientific, A11012). .. Nuclei were counterstained with DAPI (ThermoFisher Scientific, P36935).

Article Title: Loss of Reelin protects against atherosclerosis by reducing leukocyte-endothelial adhesion and lesion macrophage accumulation
Article Snippet: .. Quantitative immunostaining was performed using primary antibodies against Mac-3 (1:200; BD PharmingenTM), α-smooth muscle actin (1:200; Abcam), CD106 (1:40; BD PharmingenTM) or CD54 (1:40; R&D systems), and fluorescently-labeled secondary antibodies goat anti-rat Alexa Fluor 488 (ThermoFisher Scientific, {"type":"entrez-nucleotide","attrs":{"text":"A11006","term_id":"492389","term_text":"A11006"}} A11006 ), donkey anti-goat Alexa Fluor 594 (ThermoFisher Scientific, {"type":"entrez-nucleotide","attrs":{"text":"A11058","term_id":"489256","term_text":"A11058"}} A11058 ), or goat anti-rabbit Alexa Fluor 594 (ThermoFisher Scientific, {"type":"entrez-nucleotide","attrs":{"text":"A11012","term_id":"490206","term_text":"A11012"}} A11012 ). .. Nuclei were counterstained with DAPI (ThermoFisher Scientific, {"type":"entrez-protein","attrs":{"text":"P36935","term_id":"549826","term_text":"P36935"}} P36935 ).

Staining:

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing
Article Snippet: After 24 h of siR-β-catenin or 80 nM cisplatin treatment, the T47D and MCF-7 cells (5×10 5 ) were collected and washed twice with PBS containing 0.2% BSA (cat. no. ST023; Beyotime Institute of Biotechnology). .. The cells were stained with phycoerythrin-labeled mono-clonal CD44 (cat. no. MAB6127; 1:200; R&D Systems, Inc.) or allophycocyanin-labeled CD54 (cat. no. BBA20; 1:300; R&D Systems, Inc.) antibodies or the isotype controls (cat. no. MAB0031; 1:200; R&D Systems, Inc.) at 37°C for 30 min, rinsed twice with PBS and fixed in 10% (v/v) formaldehyde and PBS at 37°C for 25 min. Then, the cells were sorted and observed using a BD FACSCalibur 4-color flow cytometer (BD Biosciences). .. Statistical analysis was performed using FlowJo 7.6 (FlowJo LLC).

Flow Cytometry:

Blocking Assay:

Article Title: A mutation within the SH2 domain of slp-76 regulates the tissue distribution and cytokine production of iNKT cells in mice.
Article Snippet: .. Blocking antibody experiments Blocking antibodies against mouse CD11a (LFA-1; M17/4), CD54 (ICAM-1; YNI.7.4), CD11b (Mac-1; M1/70) and rat IgG2a Control (2A3) or NP-1 (761704) were obtained from BioXCell or R&D Systems. ..

Control:

Incubation:

Article Title: Ethanol, ethyl and sodium pyruvate decrease the inflammatory responses of human lung epithelial cells via Akt and NF-κB in vitro but have a low impact on hepatocellular cells.
Article Snippet: .. Thereafter, cells were incubated with a fluorescein-conjugated monoclonal antibody directed against CD54/intercellular adhesion molecule (ICAM)-1 (BBA20; R&D Systems) for 60 min at 4 ̊C as described previously (30,31). .. CD54 expression was determined by flow cytometry using a FACSCalibur (1x104 cells/scan were counted; BD Biosciences, Heidelberg, Germany) and expressed as mean fluorescence units (MFU).

Image Search Results

HeLa cells stably expressing EndoA3-GFP (green) and transiently expressing ICAM1-mScarlet (red) were imaged for 2 min using live-cell TIRF microscopy, with 1-s intervals between frames. Representative of two independent experiments. Scale bar: 10 µm.

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Figure Lengend Snippet: HeLa cells stably expressing EndoA3-GFP (green) and transiently expressing ICAM1-mScarlet (red) were imaged for 2 min using live-cell TIRF microscopy, with 1-s intervals between frames. Representative of two independent experiments. Scale bar: 10 µm.

Article Snippet: First, anti-ALCAM (Bio-Rad, MCA1926) and anti-ICAM1 (Bio-Rad, MCA1615) antibodies were labeled with benzylguanine (BG) by incubating them overnight at 4°C with a threefold molar excess of BG-GLA-NHS reagent (New England Biolabs, S9151S; prepared in anhydrous DMSO).

Techniques:

LB33-MEL cells stably expressing EndoA3-GFP (green) and transiently expressing ICAM1-mScarlet (red) were imaged for 2 min using live-cell TIRF microscopy, with 1-s intervals between frames. Representative of two independent experiments. Scale bars: 10 μm (full-size image) and 2 μm (enlarged cropped area).

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Figure Lengend Snippet: LB33-MEL cells stably expressing EndoA3-GFP (green) and transiently expressing ICAM1-mScarlet (red) were imaged for 2 min using live-cell TIRF microscopy, with 1-s intervals between frames. Representative of two independent experiments. Scale bars: 10 μm (full-size image) and 2 μm (enlarged cropped area).

Techniques:

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Figure Lengend Snippet: HeLa cells stably expressing EndoA3-GFP (green) and transiently expressing ICAM1-mScarlet (red) were imaged for 2 min using live-cell TIRF microscopy, with 1-s intervals between frames. The punctate ICAM1 structure is indicated by a white arrowhead. Representative of two independent experiments. Scale bar: 10 µm.

Techniques:

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Figure Lengend Snippet: LB33-MEL cells stably expressing EndoA3-GFP (green) and transiently expressing ICAM1-mScarlet (red) were imaged for 2 min using live-cell TIRF microscopy, with 1-s intervals between frames. The punctate ICAM1 structure is indicated by a white arrowhead. Representative of two independent experiments. Scale bars: 10 μm (full-size image) and 2 μm (enlarged cropped area).

Techniques:

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Techniques:

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Techniques:

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Figure Lengend Snippet: LB33-MEL cells stably expressing EndoA3-GFP (green) and transiently expressing ICAM1-mScarlet (red) were imaged for 2 min using live-cell TIRF microscopy, with 1-s intervals between frames. The punctate ICAM1 structure is indicated by a white arrowhead. Representative of two independent experiments. Scale bar: 10 µm.

Techniques:

A HeLa cell transiently expressing ICAM1-EGFP (green) forming an immune synapse-like conjugate with a CD8 T cell (red) was imaged by live-cell spinning-disk confocal microscopy, for 3 min, with 1-s intervals between frames. ICAM1-positive tubulo-vesicular carriers are observed fusing at the ICAM1-enriched contact zone, which grows and expands over time. Notably, anterograde transport of ICAM1-positive carriers toward the contact zone appears stronger than retrograde transport, and carrier density is higher in the region of the HeLa cell engaged in the immune synapse compared with regions not involved in synapse formation. Associated tracking and quantification data are shown in <xref ref-type=

Figure 4C–G . Representative of two independent experiments. Scale bar: 20 µm. " width="100%" height="100%">

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Figure Lengend Snippet: A HeLa cell transiently expressing ICAM1-EGFP (green) forming an immune synapse-like conjugate with a CD8 T cell (red) was imaged by live-cell spinning-disk confocal microscopy, for 3 min, with 1-s intervals between frames. ICAM1-positive tubulo-vesicular carriers are observed fusing at the ICAM1-enriched contact zone, which grows and expands over time. Notably, anterograde transport of ICAM1-positive carriers toward the contact zone appears stronger than retrograde transport, and carrier density is higher in the region of the HeLa cell engaged in the immune synapse compared with regions not involved in synapse formation. Associated tracking and quantification data are shown in Figure 4C–G . Representative of two independent experiments. Scale bar: 20 µm.

Techniques:

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Techniques:

$( A ) Illustration of the SNAP-tag-based BG-labeled antibody uptake assay to study membrane protein endocytosis and retrograde transport. ( B ) Confocal images of GalT-GFP-SNAP (green) and TMR-Star (red) in HeLa cells stably expressing the Golgi-resident GFP-fused SNAP-tag construct (HeLa GalT-GFP-SNAP). Actin (phalloidin, white) and nuclei (DAPI, blue) were also stained. Fluorescence intensity profile was made along the dashed line region in enlarged cropped area and shows the colocalization of both signals. Scale bar: 20 μm. ( C–F ) Retrograde transport of ALCAM and ICAM1. Continuous BG-labeled anti-ALCAM ( C, E ) and anti-ICAM1 ( D, F ) antibody uptake for 4 h at 37°C in HeLa GalT-GFP-SNAP cells. ( C, D ) Western blot analysis of HeLa GalT-GFP-SNAP cells transfected for 72 h with siRNAs: negative control (siCtrl) or against retromer subunits (siVPS35 and siVPS26A). Immunodetection made with anti-SNAP, anti-VPS35, anti-VPS26A, and anti-α-Tubulin (loading control) antibodies. Quantification of the covalent IgG-SNAP-GFP-GalT complex is shown as fractions of siCtrl condition (histogram). Quantification of VPS35 and VPS26A depletion is shown in . ( E, F ) Western blot analysis of HeLa GalT-GFP-SNAP cells transfected for 72 h with siRNAs: negative control (siCtrl) or against EndoA3 (siEndoA3). Immunodetection made with anti-SNAP, anti-EndoA3, and anti-clathrin heavy chain (CHC, loading control) antibodies. Quantification of the covalent IgG-SNAP-GFP-GalT complex (IB:anti-SNAP) is shown as fractions of siCtrl condition (histogram). Quantification of EndoA3 depletion is shown in . Data information: In ( B ), images are from a single experiment. Quantification data ( C–F ) are pooled from three independent experiments. Data are presented as mean ± SEM. *p<0.05, **p<0.01. One-sample t test and Wilcoxon test. Figure 1—source data 1. Original files for western blot analyses displayed in . Figure 1—source data 2. PDF files containing original western blots for .$

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Figure Lengend Snippet: ( A ) Illustration of the SNAP-tag-based BG-labeled antibody uptake assay to study membrane protein endocytosis and retrograde transport. ( B ) Confocal images of GalT-GFP-SNAP (green) and TMR-Star (red) in HeLa cells stably expressing the Golgi-resident GFP-fused SNAP-tag construct (HeLa GalT-GFP-SNAP). Actin (phalloidin, white) and nuclei (DAPI, blue) were also stained. Fluorescence intensity profile was made along the dashed line region in enlarged cropped area and shows the colocalization of both signals. Scale bar: 20 μm. ( C–F ) Retrograde transport of ALCAM and ICAM1. Continuous BG-labeled anti-ALCAM ( C, E ) and anti-ICAM1 ( D, F ) antibody uptake for 4 h at 37°C in HeLa GalT-GFP-SNAP cells. ( C, D ) Western blot analysis of HeLa GalT-GFP-SNAP cells transfected for 72 h with siRNAs: negative control (siCtrl) or against retromer subunits (siVPS35 and siVPS26A). Immunodetection made with anti-SNAP, anti-VPS35, anti-VPS26A, and anti-α-Tubulin (loading control) antibodies. Quantification of the covalent IgG-SNAP-GFP-GalT complex is shown as fractions of siCtrl condition (histogram). Quantification of VPS35 and VPS26A depletion is shown in . ( E, F ) Western blot analysis of HeLa GalT-GFP-SNAP cells transfected for 72 h with siRNAs: negative control (siCtrl) or against EndoA3 (siEndoA3). Immunodetection made with anti-SNAP, anti-EndoA3, and anti-clathrin heavy chain (CHC, loading control) antibodies. Quantification of the covalent IgG-SNAP-GFP-GalT complex (IB:anti-SNAP) is shown as fractions of siCtrl condition (histogram). Quantification of EndoA3 depletion is shown in . Data information: In ( B ), images are from a single experiment. Quantification data ( C–F ) are pooled from three independent experiments. Data are presented as mean ± SEM. *p<0.05, **p<0.01. One-sample t test and Wilcoxon test. Figure 1—source data 1. Original files for western blot analyses displayed in . Figure 1—source data 2. PDF files containing original western blots for .

Techniques: Labeling, Membrane, Stable Transfection, Expressing, Construct, Staining, Fluorescence, Western Blot, Transfection, Negative Control, Immunodetection, Control

$( A ) Confocal images of GalT-GFP-SNAP (green) and TMR-Star (red) in LB33-MEL cells stably expressing the Golgi-resident GFP-fused SNAP-tag construct (LB33-MEL GalT-GFP-SNAP). Actin (phalloidin, white) and nuclei (DAPI, blue) were also stained. The fluorescence intensity profile was made along the dashed line region in the enlarged cropped area and shows the colocalization of both signals. Scale bar: 20 μm. ( B ) Confocal images of GalT-GFP-SNAP (green) and TGN46 (red) in LB33-MEL GalT-GFP-SNAP cells. Nuclei (DAPI, blue) were also stained. The fluorescence intensity profile was made along the dashed line region in enlarged cropped area and shows the colocalization of both signals, indicating that GalT-GFP-SNAP is correctly localized at the TGN . Scale bar: 10 μm. ( C ) Quantifications of the immunoblots shown in (IB: anti-VPS35 and anti-VPS26A) confirm depletion efficiency of VPS35 and VPS26A in HeLa GalT-GFP-SNAP cells. ( D ) Retrograde transport of ICAM-1. Continuous BG-labeled anti-ICAM1 antibody uptake for 4 h at 37°C in LB33-MEL GalT-GFP-SNAP cells. Western blot analysis of LB33-MEL GalT-GFP-SNAP cells transfected with siRNAs: negative control (siCtrl) or against retromer subunits (siVPS35 and siVPS26A). Immunodetection with anti-SNAP, anti-VPS35, anti-VPS26A, and anti-α-Tubulin (loading control) antibodies. Quantification of the covalent IgG-SNAP complex is shown as fractions of siCtrl condition (histogram). Quantification of VPS35 and VPS26A depletion is shown in ( E ). ( E ) Quantifications of the immunoblots shown in ( D ) confirm depletion efficiency of VPS35 and VPS26A in LB33-MEL GalT-GFP-SNAP cells. ( F ) Retrograde transport of ALCAM. Continuous BG-labeled anti-ALCAM antibody uptake for 4 h at 37°C in HeLa GalT-GFP-SNAP cells. Western blot analysis of HeLa GalT-GFP-SNAP cells transfected for 72 h with siRNAs: negative control (siCtrl) or against Rab6 (siRab6). Immunodetection made with anti-SNAP, anti-Rab6, and anti-clathrin heavy chain (CHC, loading control) antibodies. Quantification of the covalent IgG-SNAP-GFP-GalT complex is shown as fractions of siCtrl condition (histogram). Quantification of Rab6 depletion is shown in ( G ). ( G ) Quantifications of the immunoblots shown in ( F ) confirm depletion efficiency of Rab6 in HeLa GalT-GFP-SNAP cells. ( H ) Quantifications of the immunoblots shown in (IB: anti-EndoA3) confirm depletion efficiency of EndoA3 in HeLa GalT-GFP-SNAP cells. ( I ) Retrograde transport of ALCAM. Continuous BG-labeled anti-ALCAM antibody uptake for 4 h at 37°C in LB33-MEL GalT-GFP-SNAP cells. Western blot analysis of LB33-MEL GalT-GFP-SNAP cells transfected (or not) with plasmids encoding free GFP (GFP+) or EndoA3-GFP (EndoA3+). Immunodetection with anti-SNAP, anti-EndoA3, and anti-α-Tubulin (loading control) antibodies. Quantification of the covalent IgG-SNAP-GFP-GalT complex is shown as fractions of the GFP +condition (histogram). Data information: In ( A, B ), images are from a single experiment. In ( C ), data are pooled from six independent experiments. In ( D–H ), data are pooled from three independent experiments. In ( I ), data are pooled from two independent experiments. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. One-sample t test and Wilcoxon test. Figure 1—figure supplement 1—source data 1. Original files for western blot analyses displayed in . Figure 1—figure supplement 1—source data 2. PDF files containing original western blots for .$

Journal: eLife

Article Title: Clathrin-independent endocytosis and retrograde transport in cancer cells tune immune synapse organization and CD8 T cell response

doi: 10.7554/eLife.105821

Figure Lengend Snippet: ( A ) Confocal images of GalT-GFP-SNAP (green) and TMR-Star (red) in LB33-MEL cells stably expressing the Golgi-resident GFP-fused SNAP-tag construct (LB33-MEL GalT-GFP-SNAP). Actin (phalloidin, white) and nuclei (DAPI, blue) were also stained. The fluorescence intensity profile was made along the dashed line region in the enlarged cropped area and shows the colocalization of both signals. Scale bar: 20 μm. ( B ) Confocal images of GalT-GFP-SNAP (green) and TGN46 (red) in LB33-MEL GalT-GFP-SNAP cells. Nuclei (DAPI, blue) were also stained. The fluorescence intensity profile was made along the dashed line region in enlarged cropped area and shows the colocalization of both signals, indicating that GalT-GFP-SNAP is correctly localized at the TGN . Scale bar: 10 μm. ( C ) Quantifications of the immunoblots shown in (IB: anti-VPS35 and anti-VPS26A) confirm depletion efficiency of VPS35 and VPS26A in HeLa GalT-GFP-SNAP cells. ( D ) Retrograde transport of ICAM-1. Continuous BG-labeled anti-ICAM1 antibody uptake for 4 h at 37°C in LB33-MEL GalT-GFP-SNAP cells. Western blot analysis of LB33-MEL GalT-GFP-SNAP cells transfected with siRNAs: negative control (siCtrl) or against retromer subunits (siVPS35 and siVPS26A). Immunodetection with anti-SNAP, anti-VPS35, anti-VPS26A, and anti-α-Tubulin (loading control) antibodies. Quantification of the covalent IgG-SNAP complex is shown as fractions of siCtrl condition (histogram). Quantification of VPS35 and VPS26A depletion is shown in ( E ). ( E ) Quantifications of the immunoblots shown in ( D ) confirm depletion efficiency of VPS35 and VPS26A in LB33-MEL GalT-GFP-SNAP cells. ( F ) Retrograde transport of ALCAM. Continuous BG-labeled anti-ALCAM antibody uptake for 4 h at 37°C in HeLa GalT-GFP-SNAP cells. Western blot analysis of HeLa GalT-GFP-SNAP cells transfected for 72 h with siRNAs: negative control (siCtrl) or against Rab6 (siRab6). Immunodetection made with anti-SNAP, anti-Rab6, and anti-clathrin heavy chain (CHC, loading control) antibodies. Quantification of the covalent IgG-SNAP-GFP-GalT complex is shown as fractions of siCtrl condition (histogram). Quantification of Rab6 depletion is shown in ( G ). ( G ) Quantifications of the immunoblots shown in ( F ) confirm depletion efficiency of Rab6 in HeLa GalT-GFP-SNAP cells. ( H ) Quantifications of the immunoblots shown in (IB: anti-EndoA3) confirm depletion efficiency of EndoA3 in HeLa GalT-GFP-SNAP cells. ( I ) Retrograde transport of ALCAM. Continuous BG-labeled anti-ALCAM antibody uptake for 4 h at 37°C in LB33-MEL GalT-GFP-SNAP cells. Western blot analysis of LB33-MEL GalT-GFP-SNAP cells transfected (or not) with plasmids encoding free GFP (GFP+) or EndoA3-GFP (EndoA3+). Immunodetection with anti-SNAP, anti-EndoA3, and anti-α-Tubulin (loading control) antibodies. Quantification of the covalent IgG-SNAP-GFP-GalT complex is shown as fractions of the GFP +condition (histogram). Data information: In ( A, B ), images are from a single experiment. In ( C ), data are pooled from six independent experiments. In ( D–H ), data are pooled from three independent experiments. In ( I ), data are pooled from two independent experiments. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. One-sample t test and Wilcoxon test. Figure 1—figure supplement 1—source data 1. Original files for western blot analyses displayed in . Figure 1—figure supplement 1—source data 2. PDF files containing original western blots for .

Techniques: Stable Transfection, Expressing, Construct, Staining, Fluorescence, Western Blot, Labeling, Transfection, Negative Control, Immunodetection, Control

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