Anti-GFP antibody [RM1064] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

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 ab316291 at 1/5000 (0.11 ug/ml) dilution, 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 upon images are applied with Anti-GFP antibody (ab316291) at 1/5000 dilution and the below images are applied with Anti-GFP antibody (ab290) at 1/5000 dilution.
The section was incubated with ab316291 or ab290 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 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 [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (human embryonic kidney epithelial cell) cells transfected with an empty expression vector containing a myc-His-tag® (upper left and upper right) /293T cells transfected with an GFP expression vector containing an eGFP tag (bottom left and bottom Right) cells labelling GFP with ab316291 at 1/5000 dilution (0.01 ug) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

293T cells transfected with an empty expression vector containing a myc-His-tag® were labeled with anti-Myc antibody crosslinked with Alexa Fluor®647 (670/14BP).And use Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) combined with ab316291.
Isotype are Rabbit monoclonal IgG (ab172730).
293T cells transfected with an GFP expression vector containing an eGFP tag are use Goat Anti-Rabbit IgG (Alexa Fluor® 647, ab150083) combined with ab316291.
Isotype are Rabbit monoclonal IgG (ab172730).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) transfected with empty vector containing a EGFP tag. cells labelling GFP with ab316291 at 1/500 (1.096 ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing positive staining in 293T cells transfected with empty vector containing a EGFP tag and 293T cells transfected an GFP expression vector (shown in magenta), and no staining in 293T cells transfected with empty vector containing a myc tag and 293T cells. EGFP and GFP was observed in green. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling GFP with ab316291 at 1/5000 (0.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human colon in ab316291 (Image A) and weak background stainning on the human colon in ab290 (Image B).
The image A is applied with Anti-GFP antibody (ab316291) at 1/5000 dilution and the image B is applied with Anti-GFP antibody (ab290) at 1/5000 dilution.
The section was incubated with ab316291 or ab290 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ChIP

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ChIP - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

Chromatin was prepared from Hela (human epithelial cell line from cervix adenocarcinoma) transfected with GFP Tagged human CREBBP and Hela transfected with empty vector cells according to the Abcam Dual-X-ChIP protocol.Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab316291 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 25 µl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (SYBR green approach).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling GFP with ab316291 at 1/5000 (0.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse spleen.
The section was incubated with ab316291 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling GFP with ab316291 at 1/5000 (0.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on rat spleen.
The section was incubated with ab316291 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IP

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Immunoprecipitation - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

GFP was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) transfected with a GFP protein expression vector, whole cell lysate with ab316291 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316291 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HEK-293T (human embryonic kidney epithelial cell) transfected with a GFP protein expression vector, whole cell lysate
Lane 2 : ab316291 IP in HEK-293T (human embryonic kidney epithelial cell) transfected with a GFP protein expression vector, whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of primary antibody in HEK-293T (human embryonic kidney epithelial cell) transfected with a GFP protein expression vector, whole cell lysate.

The left panel is applied with Anti-GFP antibody (ab316291) at 1/1000 dilution and the right panel is applied with Anti-GFP antibody (ab290) at 1/1000 dilution.

All lanes:

Immunoprecipitation - Anti-GFP antibody [RM1064] (<a href='/en-us/products/primary-antibodies/gfp-antibody-rm1064-ab316291'>ab316291</a>) at 1/30 dilution

All lanes:

HEK-293T (human embryonic kidney epithelial cell) transfected with a GFP protein expression vector, 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: 1s

• I-ELISA

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Indirect ELISA - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab316291 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : GFP.

Antigen concentration : 1000 ng/ml

• WB

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Western blot - Anti-GFP antibody [RM1064] - BSA and Azide free (AB316292)

This data was developed using ab316291, the same antibody clone in a different buffer formulation.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

Exposure time : Lanes 1-2 : 8 seconds; Lane 3 : 37 seconds

All lanes:

Western blot - Anti-GFP antibody [RM1064] (<a href='/en-us/products/primary-antibodies/gfp-antibody-rm1064-ab316291'>ab316291</a>) at 1/5000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

293T transfected with an empty vector containing an EGFP-myc-tag, whole cell lysate at 10 µg

Lane 3:

293T transfected with an empty vector containing a GFP-tag, whole cell lysate at 20 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: 27 kDa,15 kDa

false