Monoclonal mouse anti human icam 1 clone r6.5

Manufactured by American Type Culture Collection

Monoclonal mouse anti‐human ICAM‐1 (clone R6.5) is a laboratory product that targets the Intercellular Adhesion Molecule 1 (ICAM-1). It is a mouse monoclonal antibody specific to the human ICAM-1 protein.

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4 protocols using monoclonal mouse anti human icam 1 clone r6.5

Quantifying Leukocyte-Endothelial Interactions

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Monoclonal mouse anti‐human ICAM‐1 (clone R6.5) was from ATCC (Manassas, VA) and phycoerythrin‐labeled monoclonal mouse anti‐human ICAM‐1 (clone LB‐2) was from Santa Cruz Biotechnology (Dallas, TX). Alexa Fluor 350‐labeled goat anti‐mouse IgG was from Invitrogen (Carlsbad, CA). Nonspecific mouse IgG was from Jackson ImmunoResearch (West Grove, PA). Green Fluoresbrite® polystyrene particles (100 nm in diameter) were from Polysciences (Warrington, PA). Porous transwell inserts (1.0 µm‐pore size) were from Thermo Fisher Scientific (Waltham, MA). Human sICAM‐1 ELISA kits were from Invitrogen (Carlsbad, CA). ¹²⁵Iodine (¹²⁵I) and Iodogen pre‐coated tubes were from PerkinElmer (Waltham, MA) and Thermo Fisher Scientific (Waltham, MA), respectively. MMP‐9 Inhibitor I and MMP‐2 Inhibitor I were from EMD Millipore (Billerica, MA). Unless specified, all other reagents were from Sigma‐Aldrich (St. Louis, MO).

Manthe R.L, & Muro S. (2017). ICAM‐1‐targeted nanocarriers attenuate endothelial release of soluble ICAM‐1, an inflammatory regulator. Bioengineering & Translational Medicine, 2(1), 109-119.

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ICAM-1 Immunolabeling Protocol

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Monoclonal mouse anti-human ICAM-1 (clone R6.5) was from ATCC (Manassas, VA) and phycoerythrin-labeled monoclonal mouse anti-human ICAM-1 (clone LB-2) was from Santa Cruz Biotechnology (Dallas, TX). Alexa Fluor 350-labeled goat anti-mouse IgG was from Invitrogen (Carlsbad, CA). Non-specific mouse IgG was from Jackson ImmunoResearch (West Grove, PA). Green Fluoresbrite® polystyrene particles (100 nm in diameter) were from Polysciences (Warrington, PA). Porous transwell inserts (1.0 µm-pore size) were from Thermo Fisher Scientific (Waltham, MA). Human sICAM-1 ELISA kits were from Invitrogen (Carlsbad, CA). ¹²⁵Iodine (¹²⁵I) and Iodogen pre-coated tubes were from PerkinElmer (Waltham, MA) and Thermo Fisher Scientific (Waltham, MA), respectively. MMP-9 Inhibitor I and MMP-2 Inhibitor I were from EMD Millipore (Billerica, MA). Unless specified, all other reagents were from Sigma-Aldrich (St. Louis, MO).

Manthe R.L, & Muro S. (2017). ICAM‐1‐targeted nanocarriers attenuate endothelial release of soluble ICAM‐1, an inflammatory regulator. Bioengineering & Translational Medicine, 2(1), 109-119.

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Fluorescent Probes for Cell Imaging

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Alexa Fluor-594 cholera toxin B subunit (CTB), 10,000 MW Texas Red dextran, BODIPY-FL-C₁₂-sphingomyelin, and fluorescent secondary antibodies were from Molecular Probes (Eugene, OR). Anti-CTB and anti-caveolin-1 were from Calbiochem (La Jolla, CA). Mouse monoclonal anti-human ICAM-1 (clone R6.5) was from the American Type Culture Collection (Manassas, VA). Non-fluorescent or green Fluoresbrite 100 nm-diameter polystyrene beads were from Polysciences (Warrington, PA). Recombinant human ASM ^{32 (link)} was kindly provided by Dr. Edward Schuchman (Dept. of Genetics and Genomics Sciences, Mount Sinai School of Medicine, New York, NY). Unless otherwise noted, all other reagents were from Sigma Aldrich (St. Louis, MO).

Rappaport J., Manthe R.L., Garnacho C, & Muro S. (2015). Altered Clathrin-Independent Endocytosis in Type A Niemann-Pick Disease Cells and Rescue by ICAM-1-Targeted Enzyme Delivery. Molecular pharmaceutics, 12(5), 1366-1376.

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Quantifying ICAM-1 Expression in Endothelial Cells

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Mouse monoclonal anti-human ICAM-1 (clone R6.5) and rat monoclonal anti-mouse ICAM-1 (clone YN1) were obtained from hybridomas from American Type Culture Collection (Manassas, VA). Non-specific mouse IgG was from Jackson ImmunoResearch (West Grove, PA) and monoclonal anti-VE-cadherin was from Thermo Fisher Scientific (Waltham, MA). Texas Red-labeled secondary antibodies, Texas Red-labeled dextran (10,000 MW; lysine fixable), and Alexa Fluor 350-labeled secondary antibodies were from Invitrogen (Carlsbad, CA). Recombinant human acid sphingomyelinase (ASM) was provided by Dr. Edward Schuchman (Department of Genetics and Genomics Sciences, Mount Sinai School of Medicine, New York, NY). Fluoresbrite™ 100 nm diameter polystyrene beads were from Polysciences (Warrington, PA). Cell culture reagents were from Gibco-BRL (Grand Island, NY) or Cellgro (Manassas, VA). Porous transwell inserts (1.0 μm-diameter pore) were from Thermo Fisher Scientific (Waltham, MA). ¹²⁵I and Iodogen pre-coated tubes were from Perkin Elmer (Waltham, MA) and Thermo Fisher Scientific (Waltham, MA), respectively. MMP-9 and MMP-2 inhibitors were from EMD Millipore (Billerica, MA). Recombinant human MMP-9 (67 kDa) and MMP-2 (62 kDa) were from EMD Millipore (Billerica, MA) and Sigma-Aldrich (St. Louis, MO), respectively.

Manthe R.L., Loeck M., Bhowmick T., Solomon M, & Muro S. (2020). Intertwined Mechanisms Define Transport of Anti-ICAM Nanocarriers Across the Endothelium and Brain Delivery of a Therapeutic Enzyme. Journal of controlled release : official journal of the Controlled Release Society, 324, 181-193.

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