Journal: British Journal of Cancer

Article Title: Antibodies to CD20 and MHC class II antigen bound to B-lymphoma cells accumulate in shed cytoplasmic fragments

doi: 10.1038/sj.bjc.6602131

Figure Lengend Snippet: Immunoperoxidase staining of B-lymphoma cells after overnight incubation with anti-CD20 (1F5). Cells were deposited on cytocentrifuge slides, fixed with formaldehyde, permeabilised with saponin, and stained with a biotinylated horse anti-mouse IgG, followed by a complex of streptavidin and peroxidase. Observation was with an × 40 objective, except as noted. ( A ) Raji cells, showing prominent staining of JN spots. ( B ) RL cells, showing dark staining of apparently extracellular objects, referred to as CFs. ( C ) A lower-power photograph of RL cells (× 10), to show the general staining pattern. ( D ) RL cells stained in the absence of saponin, to show antigen that is accessible without permeabilisation. Control Abs of the same subclass produced no brown staining.

Article Snippet: Peroxidase staining utilised reagents from Vector Laboratories (Burlingame, CA, USA), namely biotinylated horse anti-mouse IgG, and a complex of streptavidin and biotinylated peroxidase.

Techniques: Immunoperoxidase Staining, Incubation, Staining, Produced

Journal:

Article Title: T Cell Apoptosis in Human Heart Allografts

doi:

Figure Lengend Snippet: Production of granzyme B and perforin during rejection. A: Granzyme B is detected in part of the lymphocytes during rejection (brown staining). Original magnification, ×150. B: Perforin is produced to the same extent as granzyme B, in part of the infiltrating cells (brown staining). Original magnification, ×150. Detection in paraffin-embedded tissue was by biotinylated horse anti-mouse antibody and HRP-conjugated streptavidin, developed with DAB.

Article Snippet: The sections were washed and incubated with biotinylated horse anti-mouse antibody (1:500; lot F0425, Vector Laboratories, Burlingame, CA) or biotinylated goat anti-rabbit antibody (1:500; lot F505, Vector) in PBS/1% BSA for 30 minutes, washed again in PBS/Tween-20, and then incubated with horseradish peroxidase (HRP)-conjugated streptavidin (1:400; lot 14022721, Boehringer Mannheim, Mannheim, Germany) in PBS/1% BSA for 30 minutes.

Techniques: Staining, Produced

Journal:

Article Title: T Cell Apoptosis in Human Heart Allografts

doi:

Figure Lengend Snippet: Regulation of apoptosis in EMBs during heart allograft rejection. A: FasL expression on apoptotic cells. Double staining for FasL (green) and TUNEL (red) demonstrated that virtually all apoptotic cells expressed FasL. Original magnification, ×280. B: FasL expression on CD4+ T cells. The majority of CD4+ T cells (green) co-expressed FasL (red, arrowheads). Original magnification, ×210. C: FasL expression on macrophages. All CD68+ macrophages (green) co-expressed FasL (red). Original magnification, ×190. D: Fas expression, shown by conventional immunoperoxidase staining (brown). Most of the infiltrating cells expressed Fas. Original magnification, ×100. E: Bax expression in apoptotic cells. Bax (green) was expressed on 50% to 70% of all apoptotic cells (TUNEL, red). Original magnification, ×330. F: Bcl2 expression in the infiltrate. Approximately 50% of all cells expressed Bcl2 (brown staining). Original magnification, ×250. For FasL, detection was by SWARPO and direct (green) or indirect (red) signal amplification. For TUNEL, detection was by TRITC-conjugated streptavidin. For CD4 and CD68, staining was with FITC-conjugated antibodies and direct signal amplification. For Bax, detection was by RAMPO and direct signal amplification. For Fas and Bcl2, detection was by biotinylated horse anti-mouse antibody and HRP-conjugated streptavidin, developed with DAB.

Article Snippet: The sections were washed and incubated with biotinylated horse anti-mouse antibody (1:500; lot F0425, Vector Laboratories, Burlingame, CA) or biotinylated goat anti-rabbit antibody (1:500; lot F505, Vector) in PBS/1% BSA for 30 minutes, washed again in PBS/Tween-20, and then incubated with horseradish peroxidase (HRP)-conjugated streptavidin (1:400; lot 14022721, Boehringer Mannheim, Mannheim, Germany) in PBS/1% BSA for 30 minutes.

Techniques: Expressing, Double Staining, TUNEL Assay, Immunoperoxidase Staining, Staining, Amplification