Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

ICC/IF image of ab89984 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 (ab89984, 10μg/ml) overnight at +4°C. The secondary antibody (orange) was ab150176 Alexa Fluor® 594 goat anti-chicken IgY (H+L) used at 2μg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Immunofluorescence staining of SOX9 using ab185230 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab185230 at 5 μg/ml, ab4674 (anti-GFAP) at 1/1000 dilution and ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 594) preadsorbed (shown in red) and ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Immunofluorescence staining of GFAP using ab278054 in primary rat hippocampal mixed glia, (prepared from P2 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDPHP4m), DIV4. The cells were fixed with 100% MeOH (5 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab278054 at 0.1 μg/ml and ab4674, Anti-GFAP antibody, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown. The antibody ab278054 gave comparable results using 4% formaldehyde fixation (10 min).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Immunofluorescence staining of GFAP using ab33922 in primary rat hippocampal mixed glia, (prepared from P2 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDPHP4m), DIV4. The cells were fixed with 100% MeOH (5 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab33922 at 0.2 μg/ml and ab4674, Anti-GFAP antibody, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown. The antibody ab33922 gave comparable results using 4% formaldehyde fixation (10 min).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Immunofluorescence staining of GFAP using ab207165 in primary rat hippocampal mixed glia, (prepared from P2 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDPHP4m), DIV4. The cells were fixed with 100% MeOH (5 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab207165 at 0.1 μg/ml and ab4674, Anti-GFAP antibody, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown. The antibody ab207165 gave comparable results using 4% formaldehyde fixation (10 min).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Immunofluorescence staining of GFAP using ab68428 in primary rat hippocampal mixed glia, (prepared from P2 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDPHP4m), DIV4. The cells were fixed with 100% MeOH (5 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab68428 at 0.1 μg/ml and ab4674, Anti-GFAP antibody, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown. The antibody ab68428 gave comparable results using 4% formaldehyde fixation (10 min).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Immunofluorescence staining of Iba-1 using ab178846 in primary rat hippocampal mixed glia (prepared from P2 rat hippocampal brain area obtained from Transnetyx Tissue by BrainBits LLC cat.no. SDPHP4m) DIV4. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab178846 at 0.1 μg/ml and ab4674 Anti-GFAP antibody at 1/1000 dilution. Cells were then incubated with ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown. The antibody ab178846 gave comparable results using MeOH fixation (100% 5 min).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Immunofluorescence staining of SOX9 using ab196450 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab196450 at 1 μg/ml (shown in green), ab4674 (anti-GFAP) at 1/1000 dilution and ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 594) preadsorbed (shown in red) and ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Immunofluorescence staining of SOX9 using ab185966 in primary hippocampal mouse neurons/glia, (obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP), DIV14. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab185966 at 1 μg/ml, ab4674 (anti-GFAP) at 1/1000 dilution and ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green), ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (shown in red) and ab150119, Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (shown in purple), all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). As expected, most GFAP positive cells are also SOX9 positive, while NeuN positive cells are SOX9 negative. SOX9 positive cells, which are not GFAP positive (e.g. asterisk) are likely neural stem cells/ oligodendrocyte precursor cells present in the culture. Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

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 (orange) was ab150176 Alexa Fluor® 594 goat anti-chicken IgY (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• WB

AbReview51452****

Western blot - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)

Gel Running Conditions : Reduced Denaturing (15% PAGE)

Detection method : Fluorescent Secondary Antibodies

All lanes:

Western blot - Anti-GFP antibody (<a href='/en-us/products/primary-antibodies/gfp-antibody-ab13970'>ab13970</a>) at 1/2000 dilution

Lane 1:

3 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate at 25 µg

Lane 2:

2 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate at 25 µg

Lane 3:

1 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate at 25 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (ab150176) at 1/5000 dilution

Predicted band size: 27 kDa

Observed band size: 25 kDa

false

Exposure time: 30s

This image is courtesy of an anonymous Abreview

• Alexa Fluor®

Unknown

Alexa Fluor® - Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (AB150176)