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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cells labeling MAP2 with ab302487 at 1/100 dilution (10.04 μg/ml), followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing cytoplasmic staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. ab183830 Anti-MAP2 rabbit monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (1 μg/mL) (Red). The Nuclear counterstain was DAPI (Blue). The negative controls are as follows : -ve control 1 : ab302487 was used as primary antibody at 1/100 dilution, followed by an150080 at 1/1000 dilution (2 μg/mL).  -ve control 2 : ab183830 at 1/200 dilution (10 μg/mL) was used as a primary antibody, followed by ab150129 at 1/1000 (2 μg/mL).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling alpha Tubulin (acetyl K40) with ab289875 at 1/100 dilution, followed by Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) (ab150129) antibody at 1/500 dilution (Green).

Confocal image showing cytoplasmic staining in HeLa cells treated with 500 ng/ml Trichostatin A for 4 hours.

Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) (ab150129) at 1/500 dilution.

• Flow Cyt

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Flow Cytometry - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Overlay histogram showing Jurkat cells stained Goat polyclonal to Ikaros (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 donkey serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Goat polyclonal to Ikaros, 0.1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 donkey anti-goat IgG (H&L) (ab150129) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was goat IgG (polyclonal) (ab37373, 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.

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cells labeling GFAP with ab302644 at 1/250 dilution (3.992 μg/ml), followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) antibody at 1/500 dilution (4 μg/mL) (Green). Confocal image showing cytoplasmic staining in mouse primary astrocytes. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. ab10062 Anti-GFAP mouse monoclonal antibody was used to counterstain tubulin at 1/50 dilution (10 μg/ml), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Red). The Nuclear counterstain was DAPI (Blue). The negative controls are as follows : -ve control 1 : ab302644 was used as primary antibody at 1/250 dilution, followed by ab150120 at 1 : 1000 dilution (2 μg/ml) 1/1000 dilution (2 μg/mL).  -ve control 2 : ab10062 at 1 : 50 dilution (10 μg/ml) was used as a primary antibody, followed by ab150129 at 1 : 500 dilution (4 μg/ml).

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cells labeling GFAP with ab302644 at 1/250 dilution (3.992 μg/ml), followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) antibody at 1/500 dilution (4 μg/mL) (Green). Confocal image showing cytoplasmic staining in rat primary astrocytes. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. ab10062 Anti-GFAP mouse monoclonal antibody was used to counterstain tubulin at 1/50 dilution (10 μg/ml), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Red). The Nuclear counterstain was DAPI (Blue). The negative controls are as follows : -ve control 1 : ab302644 was used as primary antibody at 1/250 dilution, followed by ab150120 at 1 : 1000 dilution (2 μg/ml) 1/1000 dilution (2 μg/mL).  -ve control 2 : ab10062 at 1 : 50 dilution (10 μg/ml) was used as a primary antibody, followed by ab150129 at 1 : 500 dilution (4 μg/ml).

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh) tissue labeling GFAP with ab302644 at 1/100 (9.98 ug/ml) dilution followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) at 1/1000 2 ug/mL dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488)at 1/1000 2 ug/mL dilution.

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Immunohistochemistry (Frozen sections) - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum (fresh) tissue labeling GFAP with ab302644 at 1/100 (9.98 ug/ml) dilution followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) at 1/1000 2 ug/mL dilution (Green). Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488)at 1/1000 2 ug/mL dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling alpha Tubulin (acetyl K40) with ab289875 at 1/100 dilution, followed by Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) (ab150129) antibody at 1/500 dilution (Green).

Confocal image showing cytoplasmic staining in NIH/3T3 cells treated with 500 ng/ml Trichostatin A for 4 hours.

Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) (ab150129) at 1/500 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling non-muscle Myosin IIB/MYH10 with ab300649 at 1/50 (19.56 ug/ml) dilution, followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) antibody at 1/500 4ug/ml dilution (Green). Confocal image showing cytoskeletal staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) at 1/500 4ug/ml dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cells labeling MAP2 with ab302487 at 1/100 dilution (10.04 μg/ml), followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing cytoplasmic staining in rat primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. ab183830 Anti-MAP2 rabbit monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (1 μg/mL) (Red). The Nuclear counterstain was DAPI (Blue). The negative controls are as follows : -ve control 1 : ab302487 was used as primary antibody at 1/100 dilution, followed by ab150080 at 1/1000 dilution (2 μg/mL).  -ve control 2 : ab183830 at 1/200 dilution (10 μg/mL) was used as a primary antibody, followed by ab150129 at 1/1000 (2 μg/mL).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C6 cells labelling alpha Tubulin (acetyl K40) with ab289875 at 1/100 dilution, followed by Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) (ab150129) antibody at 1/500 dilution (Green).

Confocal image showing cytoplasmic staining in C6 cells treated with 500 ng/ml Trichostatin A for 4 hours.

Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (Alexa Fluor^® 488) (ab150129) at 1/500 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)

ICC/IF image of 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 / 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 (ab7291, 1μg/ml) overnight at +4°C. ab98800, goat anti-mouse IgG, was then added as a secondary bridging antibody, at 1/250 dilution for 1h. ab150129 Alexa Fluor® 488 donkey anti-goat IgG (H+L) (shown in green) was then used at 1μg/ml for 1h as a tertiary antibody. 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.

• Alexa Fluor®

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Alexa Fluor® - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (AB150129)