Anti-GFP antibody [3H9] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (AB255886)

This data was developed using ab252881 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a GFP construct (A) labeling GFP with ab252881 at 1/2000 dilution (0.539 μg/ml) followed by a ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). No staining observed on HEK-293T cells (B). The section was incubated with ab252881 for 30 mins at room temperature. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (AB255886)

This data was developed using ab252881 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human tonsil section labeling GFP with ab252881 at 1/2000 dilution (0.539 μg/ml) followed by a ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). The section was incubated with ab252881 for 30 mins at room temperature. Counterstained with Hematoxylin.

Negative control : no staining on human tonsil.

Secondary antibody only control : Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [3H9] - BSA and Azide free (AB255886)

This data was developed using ab252881 the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells stained for GFP (red) using ab252881 at 1/100 dilution (10.78 μg/ml), followed by ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor^® 594) antibody at 1/1000 dilution. Confocal image showing cytoplasmic and nuclear staining in HEK-293T cells transfected with GFP only plasmid. The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (AB255886)

This data was developed using ab252881 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat liver section labeling GFP with ab252881 at 1/2000 dilution (0.539 μg/ml) followed by a ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). The section was incubated with ab252881 for 30 mins at room temperature. Counterstained with Hematoxylin.

Negative control : no staining on rat liver.

Secondary antibody only control : Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [3H9] - BSA and Azide free (AB255886)

This data was developed using ab252881 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse spleen section labeling GFP with ab252881 at 1/2000 dilution (0.539 μg/ml) followed by a ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). The section was incubated with ab252881 for 30 mins at room temperature. Counterstained with Hematoxylin.

Negative control : no staining on mouse spleen.

Secondary antibody only control : Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Supplier Data

Western blot - Anti-GFP antibody [3H9] - BSA and Azide free (AB255886)

This data was developed using ab252881 the same antibody clone in a different buffer formulation.

Blocking and Diluting buffer : 5% NFDM/TBST

Exposure time : 3 seconds

All lanes:

Western blot - Anti-GFP antibody [3H9] (<a href='/en-us/products/primary-antibodies/gfp-antibody-3h9-ab252881'>ab252881</a>) at 1/5000 dilution

Lane 1:

HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with GFP expression vector, whole cell lysate at 10 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

Lane 4:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/100000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-GFP antibody [3H9] - BSA and Azide free (AB255886)

This data was developed using ab252881, the same antibody clone in a different buffer formulation.

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

ab252881 does not recognize RFP

All lanes:

Western blot - Anti-GFP antibody [3H9] (<a href='/en-us/products/primary-antibodies/gfp-antibody-3h9-ab252881'>ab252881</a>) 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

Lane 3:

293T cells transfected with RFP protein whole cell lysate 20ug at 20 µg

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/10000 dilution

Observed band size: 50 kDa

false

Exposure time: 1s