Goat Anti-Mouse IgG H&L (Alexa Fluor® 594)

3 product images

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  Flow Cytometry - Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150116)

  Overlay histogram showing Jurkat cells stained with Anti-CD3 antibody [MEM-57] ab8090 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (Anti-CD3 antibody [MEM-57] ab8090, 0.01μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 594) (ab150116) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (Mouse IgG2a, Kappa Monoclonal [B12/8] - Isotype Control ab91361, 0.01μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 561nm laser and 610/20 bandpass filter.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150116)

  ICC/IF image of Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 1μg/ml) overnight at +4°C. The secondary antibody (orange) was ab150116 Alexa Fluor^® 594 goat anti-mouse IgG (H+L) used at 2μg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.
  

  The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

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  Alexa Fluor® - Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150116)