Anti-GFP antibody [EPR28018-38]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR28018-38] (AB314656)

Immunohistochemical analysis of paraffin-embedded (A) 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a GFP-tag (B) 293T transfected with an empty vector containing an EGFP-myc-tag (C) 293T transfected with an empty vector containing a myc-His-tag® tissue labeling GFP with ab314656 at 1/4000 (0.127 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on 293T transfected with an empty vector containing a GFP-tag cell pellets (A) and 293T transfected with an empty vector containing an EGFP-myc-tag cell pellets (B). No staining on 293T transfected with an empty vector containing a myc-His-tag cell pellets (C). The section was incubated with ab314656 for 30 mins at room temperature. 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR28018-38] (AB314656)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling GFP with ab314656 at 1/50 (10.18 ug/ml) dilution followed by ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150088) antibody at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing positive staining in 293T cells transfected with a empty expression vector containing a GFP tag. Image was taken with a confocal microscope(Leica-Microsystems TCS SP8). The 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 (2 ug/mL) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR28018-38] (AB314656)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling GFP with ab314656 at 1/4000 (0.127 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human spleen. The section was incubated with ab314656 for 30 mins at room temperature. 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-GFP antibody [EPR28018-38] (AB314656)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a EGFP expression vector containing a EGFP tag (Right) cells labelling GFP with ab314656 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal isotype control - Alexa Fluor® 647 / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR28018-38] (AB314656)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling GFP with ab314656 at 1/4000 (0.127 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on rat spleen. The section was incubated with ab314656 for 30 mins at room temperature. 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR28018-38] (AB314656)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling GFP with ab314656 at 1/4000 (0.127 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on mouse spleen. The section was incubated with ab314656 for 30 mins at room temperature. 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

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Immunoprecipitation - Anti-GFP antibody [EPR28018-38] (AB314656)

GFP was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a GFP-tag, whole cell lysate with ab314656 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314656 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a GFP-tag, whole cell lysate

Lane 2 : ab314656 IP in 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a GFP-tag, whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314656 in 293T transfected with an empty vector containing a GFP-tag, whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-GFP antibody [EPR28018-38] (ab314656) at 1/30 dilution

All lanes:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a GFP-tag, whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 8s

• IP

Supplier Data

Immunoprecipitation - Anti-GFP antibody [EPR28018-38] (AB314656)

GFP was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing an EGFP-myc-tag, whole cell lysate with ab314656 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314656 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing an EGFP-myc-tag, whole cell lysate

Lane 2 : ab314656 IP in 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing an EGFP-myc-tag, whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314656 in 293T transfected with an empty vector containing an EGFP-myc-tag, whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-GFP antibody [EPR28018-38] (ab314656) at 1/30 dilution

All lanes:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing an EGFP-myc-tag, whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 26 kDa

false

Exposure time: 6s

• WB

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Western blot - Anti-GFP antibody [EPR28018-38] (AB314656)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-GFP antibody [EPR14104] (ab183734) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-GFP antibody [EPR28018-38] (ab314656) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 7 µg

Lane 2:

293T transfected with an empty vector containing an EGFP-myc-tag, whole cell lysate at 7 µg

Lane 3:

293T transfected with an empty vector containing a GFP-tag, whole cell lysate at 7 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 26 kDa,36 kDa

false

Exposure time: 3s