Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150085)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) cells labeling SIGLEC5 and SIGLEC14 with ab307846 at 1/50 dilution (9.72 µg/ml) followed by ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150088) antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing positive staining in HEK-293T cells transfected with a human SIGLEC14 or SIGLEC5 expression vector containing a myc tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150088) at 1/1000 dilution (2 µg/ml).

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150085)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh) tissue labeling Heparan Sulfate Proteoglycan 2/Perlecan with ab315029 at 1/500 (0.988 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing positive staining on mouse kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab315029 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150085)

ICC/IF image of ab8227 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab8227, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab150085 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.

• Flow Cyt

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Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150085)

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, pre-adsorbed) (ab150085) was used at 1/2000 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150085)

HeLa cells showing negative staining by ICC/IF using only secondary antibody. The cells were 4% formaldehyde fixed (10 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 secondary antibody (green) was ab150085 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.

• Alexa Fluor®

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Alexa Fluor® - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150085)