Imaging Mass Cytometry™ (IMC™) image of human lung cancer tissue stained with Anti-pan Cytokeratin antibody [AE1/AE3]. ab80826 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.

Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human colon carcinoma tissue with ab80826.

ICC/IF image of ab80826 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab80826, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

This image was generated using the ascites version of the product.

Overlay histogram showing A431 cells stained with ab80826 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab80826, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This image was generated using the ascites version of the product.

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue with ab80826 at 1/400 dilution. Anti-Mouse HRP polymer was used as the secondary detection system. Heat-mediated antigen retrieval was performed using citrate based pH 6.0 buffer.

This image was generated using the ascites version of the product.

Immunohistochemistry analysis of formalin-fixed, paraffin-embedded Human skin tissue using 1/50 ab80826, a peroxidase-conjugated secondary antibody and an AEC chromogen. Note cytoplasmic staining of epithelial cells.

This image was generated using the ascites version of the product.

Immunohistochemical analysis of rat bladder tissue, labeling pan Cytokeratin with ab80826. Samples were formaldehyde fixed, heat mediated antigen retrieval was perfmormed with 10mM citrate buffer, and blocking was with 10% serum for 2 hours at 23°C. Incubation with ab80826 (diluted 1/1000) was for 17 hours at 4°C.

This image was generated using the ascites version of the product.

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