Goat Anti-Mouse IgG H&L (Alexa Fluor® 647)

5 product images

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  Flow Cytometry - Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (ab150115)

  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, 0.1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 647) (ab150115) was used at 1/2000 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, 0.1μg/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 solid-state 25mW red diode laser (635nm) and 675/30 bandpass filter.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (ab150115)

  ICC/IF image of Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 stained HeLa cells. The cells were 4% paraformaldehyde fixed (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal donkey 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 primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 5μg/ml) overnight at +4°C. The secondary antibody (red) was ab150115 Alexa Fluor^® 647 goat anti-mouse IgG (H+L) used at 1μg/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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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (ab150115)

  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 (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 1μg/ml) and (Anti-Lamin B1 antibody - Nuclear Envelope Marker ab16048, 1μg/ml) overnight at +4°C. The secondary antibodies were ab150115 Alexa Fluor^® 647 (red) goat anti-mouse IgG (H+L) used at 2μg/ml for 1h and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Alexa Fluor^® 488 (green) goat anti-rabbit IgG (H+L) used at 2μg/ml for 1h. DAPI was used to stain the cell nuclei.

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  Alexa Fluor® - Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (ab150115)
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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (ab150115)

  Immunocytochemistry/Immunofluorescence analysis of A549 (human lung carcinoma epithelial) cells with Anti-ADX antibody [EPR4629] ab108257 at a 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Mouse anti-COX IV monoclonal antibody - Mitochondrial Marker was used as a counterstain at a 1/50 dilution followed by ab150115 Alexa Fluor^{®,/sup> 647 Goat anti-Mouse secondary antibody used at a 1/1000 dilution.}

  The negative controls are shown in bottom middle and right hand panels - for negative control 1, Anti-ADX antibody [EPR4629] ab108257 was used at a dilution of 1/100 followed by Alexa Fluor^® 647 Goat anti-Mouse secondary ab150115 at a dilution of 1/1000. For negative control 2, Mouse anti-COX IV monoclonal antibody - Mitochondrial Marker was used at a dilution of 1/50 followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a dilution of 1/1000.

  Confocal image showing cytoplasmic staining in A549 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).