Anti-T7 tag® antibody [EPR28033-8]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-T7 tag® antibody [EPR28033-8] (AB317258)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) transfected with a T7 tag®-CTCF expression vector cells labelling T7 tag® with ab317258 at 1/1000 (0.527 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear staining in HeLa cells transfected with a T7 tag®-CTCF expression vector (shown in green). The counterstain was observed in red. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T7 tag® antibody [EPR28033-8] (AB317258)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling T7 tag® with ab317258 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human cerebrum.The section was incubated with ab317258 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T7 tag® antibody [EPR28033-8] (AB317258)

Immunohistochemical analysis of paraffin-embedded (A) Hela (human cervix adenocarcinoma epithelial cell) transfected with a T7 tag®-CTCF expression vector cell pellet. (B) Hela transfected with empty vector cell pellet tissue labeling T7 tag® with ab317258 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Hela transfected with a T7 tag®-CTCF expression vector cell pellet (image A). No staining on Hela transfected with empty vector cell pellet (image B).The section was incubated with ab317258 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-T7 tag® antibody [EPR28033-8] (AB317258)

T7 tag® was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) transfected with an Human CTCF expression vector containing a T7 tag® whole cell lysate with ab317258 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317258 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) transfected with an Human CTCF expression vector containing a T7 tag®, whole cell lysate
Lane 2 : ab317258 IP in HeLa (human cervix adenocarcinoma epithelial cell) transfected with an Human CTCF expression vector containing a T7 tag® whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317258 in HeLa transfected with an Human CTCF expression vector containing a T7 tag® whole cell lysate

All lanes:

Immunoprecipitation - Anti-T7 tag® antibody [EPR28033-8] (ab317258) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with an Human CTCF expression vector containing a T7 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: 41s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T7 tag® antibody [EPR28033-8] (AB317258)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling T7 tag® with ab317258 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat cerebrum.The section was incubated with ab317258 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-T7 tag® antibody [EPR28033-8] (AB317258)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling T7 tag® with ab317258 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse cerebrum.The section was incubated with ab317258 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

• WB

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Western blot - Anti-T7 tag® antibody [EPR28033-8] (AB317258)

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

In Western blot, Anti-CTCF antibody [EPR7314 (B)] - ChIP Grade (ab128873) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-T7 tag® antibody [EPR28033-8] (ab317258) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with empty vector, whole cell lysate at 20 µg

Lane 2:

HeLa transfected with an Human CTCF expression vector containi a T7 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: 150 kDa,36 kDa

false

Exposure time: 8s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-T7 tag® antibody [EPR28033-8] (AB317258)

Indirect ELISA analysis of ab317258 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 : T7 tag®.

Antigen concentration : 1000 ng/ml

• WB

Supplier Data

Western blot - Anti-T7 tag® antibody [EPR28033-8] (AB317258)

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-T7 tag® antibody [EPR28033-8] (ab317258) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an Human Histone H3.3 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) transfected with an Human Histone H3 expression vector containing a T7 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: 15 kDa

false

Exposure time: 55s