Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Overlay histogram showing Jurkat cells stained with ab16669 (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 (ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (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 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Flow cytometric analysis of HEK-293T (Human embryonic kidney epithelial cell, Left panel) / Caco-2 (Human colorectal adenocarcinoma epithelial cell, Right panel) using ab278053 at 1/500 dilution (0.1µg) (red). The isotype control used was the Rabbit monoclonal IgG (ab172730), black line and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Secondary antibody used was the Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution.
Negative control: HEK-293T. (PMID 20167130)
Gated on viable cells.
Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
ICC/IF image of beta Tubulin staining in 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 primary antibody (ab6046, 5μg/ml) overnight at +4°C. The secondary antibody (green) was ab150077 Alexa Fluor® 488 goat anti-rabbit 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.
Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Immunocytochemistry/ Immunofluorescence analysis of C6(Rat glial tumor glial cell) cells labeling alpha smooth muscle Actin with purified ab32575 at 1/100 dilution (0.71 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Immunocytochemistry analysis of SK-OV-3 (human ovarian cancer epithelial cell)cells labelling Zic2 with ab150404 at 2.5 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).
Immunohistochemistry (Frozen sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Immunohistochemistry (Frozen sections) analysis of rat stomach tissue (4% PFA, 0.2% Tritron x-100) labelling Integrin alpha 6 with ab240252 at 1/100 dilution. ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. Negative control using PBS instead of primary antibody (left). Counterstained with DAPI.
Positive staining was seen on rat stomach.
