Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

Immunohistochemical analysis of paraffin-embedded human tonsil labeling GFP with ab300643 at 1/10000 dilution (0.996 µg/ml) followed by ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. No staining on human tonsil. The section was incubated with ab300643 for 30 mins at room temperature followed by anti-chicken IgG antibody (ab97136) for 8 mins during the Leica DS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

Flow Cytometry (Intracellular) analysis of 293T transfected with GFP tagged Firefly Luciferase over expression vector labeling GFP (right) with ab300643 at a 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti-Chicken IgY H&L (Alexa Fluor® 647, ab150175) was used as the secondary antibody at a 1/5000 dilution.  Chicken IgY Isotype control. (left)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T transfected with GFP expression vector containing a his tag. (B) Wild type HEK-293T cells labeling GFP with ab300643 at 1/10000 dilution (0.996 µg/ml) followed by ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on HEK-293T transfected with GFP expression vector containing a his tag (Image : A); No staining on Wild type HEK-293T cells (Image : B). The section was incubated with ab300643 for 30 mins at room temperature followed by anti-chicken IgG antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized HEK293T (human embryonic kidney epithelial cells) labeling GFP with ab300643 at 1/50 dilution followed by ab150175 Goat Anti-Chicken IgY H&L (Alexa Fluor® 647) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (red). Confocal image showing positive staining in HEK-293T cells transfected with a Firefly Luciferase expression vector containing a GFP tag. The nuclear counter stain is DAPI (blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized 293/GFP (human embryonic kidney epithelial cell) cells labeling GFP with ab300643 at 1/50 dilution followed by ab150175 Goat Anti-Chicken IgY H&L (Alexa Fluor® 647) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (red). Confocal image showing positive staining in 293/GFP cells. The nuclear counter stain is DAPI (blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

Immunohistochemical analysis of paraffin-embedded mouse spleen labeling GFP with ab300643 at 1/10000 dilution (0.996 µg/ml) followed by ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. No staining on mouse spleen. The section was incubated with ab300643 for 30 mins at room temperature followed by anti-chicken IgG antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

Immunohistochemical analysis of paraffin-embedded rat spleen labeling GFP with ab300643 at 1/10000 dilution (0.996 µg/ml) followed by ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. No staining on rat spleen. The section was incubated with ab300643 for 30 mins at room temperature followed by anti-chicken IgG antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• WB

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Western blot - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

In Western blot, anti-GAPDH antibody (ab181602) loading control staining.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T cells transfected with an empty vector whole cell lysate at 20 µg

Lanes 3 - 4:

293T cells transfected with GFP tagged IFNA2 protein whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/20000 dilution

Observed band size: 50 kDa

false

Exposure time: 40s

• WB

Supplier Data

Western blot - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (AB300643)

Blocking / Diluting buffer and concentration : 5% NFDM/TBST Exposure time : 3.25 seconds

All lanes:

Western blot - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643) at 1/1000 dilution

Lane 1:

Non-transfected 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

GFP transfected 293 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/20000 dilution

Observed band size: 27 kDa

false