Anti-GFP 抗体 [EPR14104]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (AB183734)

Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse liver tissue (left) and normal mouse liver tissue (right) labeling GFP with unpurified ab183734 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

AbReview76836****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (AB183734)

Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed, mouse pineal gland tissue from an adult Cx3cr1-eGFP+/-; APP/PS1 mouse. Stained with ab183734 at 1/300 dilution. Secondary antibody used was Alexa Fluor® 647 donkey Anti-rabbit antibody at 1/1000 dilution. Cell nuclei were counterstained with DAPI (grey). Blocking was done with 3% BSA for 1 hour at 24°C. The sample was incubated with the primary antibody and PBS with 3% BSA and 1% Triton X-100 for 16 hours at 24°C. Antigen retrieval method was heat mediated 10 mM citrate buffer pH6

This image is courtesy of an Abreview submitted by Estela Munoz

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (AB183734)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse pancreas tissue (negative control) labelling GFP with purified ab183734 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] (AB183734)

Immunofluorescent analysis of 4% paraformaldehyde-fixed GFP transfected 293 cells labeling GFP with unpurified ab183734 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody at 1/200 dilution (green). Counterstained with Dapi (blue)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] (AB183734)

ab183734 staining GFP in GFP-transfected NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab183734 at 1/500 dilution overnight at +4°C followed by incubation with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488), for 1 hour, at 1μg/ml.

Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH3T3 cells, the cells upon which ab183734 was applied gave a stronger signal in the 488 channel, indicating that ab183734 is binding to GFP and therefore eliciting signal amplification.

ab183734 was also applied to non-GFP-transfected NIH3T3 cells, which produced no positive staining, indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (AB183734)

Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse colon tissue (left) and normal mouse colon tissue (right) labeling GFP with unpurified ab183734 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-GFP antibody [EPR14104] (AB183734)

Intracellular Flow Cytometry analysis of 293T (human embryonic kidney) transfected with human TNFRSF9 cells labeling GFP with purified ab183734 at 1/50 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 647) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] (AB183734)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of GFP transgenic mouse pancreas tissue labelling GFP with purified ab183734 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

Lab

Western blot - Anti-GFP antibody [EPR14104] (AB183734)

Different batches of ab183734 were tested on GFP transfected HEK-293 (Human embryonic kidney epithelial cell) lysate at 0.004 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 27 kDa.

All lanes:

Western blot - Anti-GFP antibody [EPR14104] (ab183734)

Predicted band size: 27 kDa

false

• WB

Lab

Western blot - Anti-GFP antibody [EPR14104] (AB183734)

Blocking and dilution buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-GFP antibody [EPR14104] (ab183734) at 1/10000 dilution

Lane 1:

GFP transfected 293 cell lysate at 10 µg

Lane 2:

Non-transfected 293 cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 27 kDa

Observed band size: 25 kDa

false

• WB

Supplier Data

Western blot - Anti-GFP antibody [EPR14104] (AB183734)

All lanes:

Western blot - Anti-GFP antibody [EPR28018-38] (<a href='/products/primary-antibodies/gfp-antibody-epr28018-38-ab314656'>ab314656</a>) 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='/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

• WB

Supplier Data

Western blot - Anti-GFP antibody [EPR14104] (AB183734)

All lanes:

Western blot - Anti-GFP antibody [EPR14104] (ab183734) at 1/10000 dilution

Lane 1:

GFP transfected 293 cell lysate at 10 µg

Lane 2:

Non-transfected 293 cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false