Anti-RED1 antibody [EPR25433-101]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RED1 antibody [EPR25433-101] (AB290638)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling RED1 with ab290638 at 1/500 (1.206 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Nuclear staining on human kidney. The section was incubated with ab290638 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RED1 antibody [EPR25433-101] (AB290638)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling RED1 with ab290638 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RED1 antibody [EPR25433-101] (AB290638)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling RED1 with ab290638 at 1/500 (1.206 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Nuclear staining on human cerebrum. The section was incubated with ab290638 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RED1 antibody [EPR25433-101] (AB290638)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling RED1 with ab290638 at 1/50 (12.06 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2μg/mL) dilution (Green). Confocal image showing nuclear staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5μg/ml) dilution (Red). 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 (2μg/mL) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RED1 antibody [EPR25433-101] (AB290638)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling RED1 with ab290638 at 1/500 (1.206 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ C Polymer Refine Detection) . Nuclear staining on rat cerebrum. The section was incubated with ab290638 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 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-RED1 antibody [EPR25433-101] (AB290638)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling RED1 with ab290638 at 1/500 (1.206 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Nuclear staining on mouse cerebrum. The section was incubated with ab290638 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RED1 antibody [EPR25433-101] (AB290638)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast adherent) cells labelling RED1 with ab290638 at 1/50 (12.06 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2μg/mL) dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5μg/ml) dilution (Red). 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 (2μg/mL) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-RED1 antibody [EPR25433-101] (AB290638)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling RED1 with ab290638 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-RED1 antibody [EPR25433-101] (AB290638)

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

All lanes:

Western blot - Anti-RED1 antibody [EPR25433-101] (ab290638) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HEL (human erythroleukemia erythroblast) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

C2C12 ( mouse myoblasts myoblast) 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

Predicted band size: 81 kDa

Observed band size: 90 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-RED1 antibody [EPR25433-101] (AB290638)

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

The bolts of lanes 1&2 and lanes 3-5 were incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG (Merck DC03L, 1 : 2000) and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051, 1 : 100, 000), respectively.

All lanes:

Western blot - Anti-RED1 antibody [EPR25433-101] (ab290638) at 1/1000 dilution

Lane 1:

Human placenta tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat placenta tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

oat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG (Merck DC03L) at 1/2000 dilution

Lanes 3 - 5:

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

Predicted band size: 81 kDa

Observed band size: 90 kDa

false

Exposure time: 48s

• WB

Unknown

Western blot - Anti-RED1 antibody [EPR25433-101] (AB290638)

This data was developed using ab290638, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-RED1 antibody [EPR25433-101] (ab290638) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

HEK-293 transfected with ADARB1(WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

HEK-293 transfected with ADARB2 (WT) expression vector containing a myc-His-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

Predicted band size: 81 kDa

Observed band size: 90 kDa

false

Exposure time: 3s