biotin conjugated rat anti mouse cd44  (Cedarlane)

Journal: Scientific Reports

Article Title: High-resolution imaging of living mammalian cells bound by nanobeads-connected antibodies in a medium using scanning electron-assisted dielectric microscopy

doi: 10.1038/srep43025

Figure Lengend Snippet: ( a ) A schematic diagram of the SE-ADM system based on FE-SEM. The liquid-sample holder with nanoparticles and/or cultured cells is mounted on the pre-amplifier-attached stage, which is introduced into the specimen chamber. The scanning electron beam is applied to the W-coated SiN film at a low acceleration voltage. The measurement terminal under the holder detects the electrical signal through liquid specimens. ( b ) Overview of the liquid holder in the cultured cancer cells bound with 100-nm beads. After 4–5 days of culturing in the dish holder, the cancer cells stained with streptavidin-conjugated 100-nm beads and biotin-conjugated anti-CD44 antibodies were sealed in the bottom sample holder. The cancer cells were attached to the upper SiN film, and its W-coated side was irradiated by the scanning electron beam. ( c ) A conceptual diagram of the cell membrane with streptavidin-conjugated 100-nm polystyrene beads and biotin-conjugated anti-CD44 antibodies via streptavidin-biotin interaction. Biotin is not shown in the diagram.

Article Snippet: Biotin-conjugated rat anti-mouse-CD44 antibodies (catalog #: 553132) were obtained from BD Bioscience.

Techniques: Cell Culture, Staining, Irradiation

Journal: Scientific Reports

Article Title: High-resolution imaging of living mammalian cells bound by nanobeads-connected antibodies in a medium using scanning electron-assisted dielectric microscopy

doi: 10.1038/srep43025

Figure Lengend Snippet: ( a ) A dielectric image of the 100-nm polystyrene beads conjugated with streptavidin in liquid at 50,000× magnification under a 3.6-kV electron beam acceleration. The image was filtered using 2D GF (11 × 11 pixels, 1.2σ) after background subtraction. Several clear black spheres dispersed over the whole area represent the 100-nm polystyrene beads. A schematic figure in the upper-right square shows polystyrene beads conjugated with streptavidin on their surface. ( b ) A dielectric image of 100-nm polystyrene beads conjugated with streptavidin bound to biotin-conjugated anti-CD44 antibodies in liquid at 50,000× magnification. The image of antibody-binding beads shows a rough surface with a small projection form. A schematic figure in the upper-right square shows polystyrene beads conjugated with streptavidin and antibodies on their surface. ( c , d ) Expanded pseudo-colour images of 100-nm beads indicated by the red arrows in ( a ). These beads surfaces are smooth. ( e , f ) Expanded pseudo-colour images of 100-nm beads to which anti-CD44 antibodies are bound (indicated by the red arrows in ( b )). These beads have very rough surfaces. ( g ) A 3D colour map of the same beads in ( c ). ( h ) A 3D colour map of the same beads to which antibodies are bound in ( e ). The white arrow suggests the anti-CD44 antibody. ( i ) Comparison of the line plots of bead centres. The black line shows the 100-nm beads of ( d ), while the red line shows the beads to which antibodies are bound in ( f ). The range of the red line is wider than that of the black line. Scale bars are 100 nm in ( a ) and ( b ), and 50 nm in ( c ) and ( e ).

Article Snippet: Biotin-conjugated rat anti-mouse-CD44 antibodies (catalog #: 553132) were obtained from BD Bioscience.

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: High-resolution imaging of living mammalian cells bound by nanobeads-connected antibodies in a medium using scanning electron-assisted dielectric microscopy

doi: 10.1038/srep43025

Figure Lengend Snippet: ( a ) Optical phase-contrast image of antibody-stained cultured cells obtained with an optical microscope at 400× magnification. ( b ) Fluorescence image of anti-CD44 immunostained cells obtained from an optical microscope with a fluorescence filter at 400× magnification. Anti-CD44 antibodies are localized on the cell membranes. The scale bar represents 10 μm in ( a ).

Article Snippet: Biotin-conjugated rat anti-mouse-CD44 antibodies (catalog #: 553132) were obtained from BD Bioscience.

Techniques: Staining, Cell Culture, Microscopy, Fluorescence

Journal: Scientific Reports

Article Title: High-resolution imaging of living mammalian cells bound by nanobeads-connected antibodies in a medium using scanning electron-assisted dielectric microscopy

doi: 10.1038/srep43025

Figure Lengend Snippet: ( a ) An image of the streptavidin-conjugated 100-nm beads binding to cells via biotin-conjugated anti-CD44 antibodies in medium using the SE-ADM system at an electron beam-acceleration of 6 kV, 5,000× magnification and −32 V bias. ( b ) An image of the streptavidin-conjugated 100-nm beads binding cells without anti-CD44 antibodies using SE-ADM system at an electron beam-acceleration of 10 kV and 3,000× magnification. ( c ) An image of unstained cancer cells at electron beam-acceleration of 8 kV and 5,000× magnification. ( d – f ) Expanded images of the red boxes in ( a – c ) with 20,000× magnification. In ( d ), many clear black spherical particles are dispersed over the whole area. In ( e ), few spherical beads are observed. In ( f ), almost no spherical beads are observed. ( g ) The average number of nano-beads/field with or without anti-CD44 antibody to the 4T1E/M3 cells. The average number of nano-beads/field is 29.5 with antibodies, 3.75 without antibodies and 1.25 in the control, in four scanned images of each condition at 20,000× magnification (an image size of 5.8 μm × 4.8 μm). Values are means ± SD; *** p < 0.0001. The scale bars represent 1 μm in ( a – c ), 500 nm in ( d – f ).

Article Snippet: Biotin-conjugated rat anti-mouse-CD44 antibodies (catalog #: 553132) were obtained from BD Bioscience.

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: High-resolution imaging of living mammalian cells bound by nanobeads-connected antibodies in a medium using scanning electron-assisted dielectric microscopy

doi: 10.1038/srep43025

Figure Lengend Snippet: ( a ) A dielectric image of the nucleus region of a cancer cell stained with antibody-binding 100-nm beads at an electron beam acceleration of 6 kV, 3,000× magnifications and −32 V bias. ( b ) An expanded image of the red boxed area in ( a ) at 10,000× magnification. ( c ) High-resolution image of the red boxed area in ( b ) at 30,000× magnification. Clear black spherical particles are dispersed over the whole area. ( d ) Another image of the cancer cells stained with antibody-binding 100-nm beads at an electron beam acceleration of 6 kV, 3,000× magnification and −32 V bias. ( e ) An expanded image of the red boxed area at the bottom centre in ( d ) at 20,000× magnification. ( f ) High-resolution image of the red boxed area in ( d ) at the top left at 40,000× magnification. This area also shows 100-nm beads of clear black spheres. ( g , h ) Pseudo-colour maps of enlarged images of the bead binding areas indicated by red arrows in ( f ). ( i ) 3D colour map of ( h ). The white arrows suggest the anti-CD44 antibody. The scale bars are 2 μm in ( a ) and ( d ), 1 μm in ( b ), 500 nm in ( e ), 200 nm in ( c ) and ( f ), 50 nm in ( g ) and ( i ).

Article Snippet: Biotin-conjugated rat anti-mouse-CD44 antibodies (catalog #: 553132) were obtained from BD Bioscience.

Techniques: Staining, Binding Assay