Anti-CUX1+CUX2 antibody [EPR26509-154]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of paraffin-embedded Rat small intestine tissue labeling CUX1+CUX2 with ab309139 at 1/500 (4.78 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Low expression tissue : almost no staining on rat small intestine. The section was incubated with ab309139 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh) tissue labeling CUX1+CUX2 with ab309139 at 1/50 (9.56 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on rat liver (PMID : 8798433). The nuclear counterstain was DAPI (Blue). The section was incubated with ab309139 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of paraffin-embedded Mouse small intestin tissue labeling CUX1+CUX2 with ab309139 at 1/500 (4.78 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Low expression tissue : almost no staining on mouse small intestine (PMID : 8879483). The section was incubated with ab309139 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh) tissue labeling CUX1+CUX2 with ab309139 at 1/50 (9.56 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on mouse liver (PMID : 8798433). The nuclear counterstain was DAPI (Blue). The section was incubated with ab309139 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CUX1+CUX2 with ab309139 at 1/500 (4.78 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse cerebrum with intense staining on superficial cortical layers. (A) Low‑powered (magnification, x40) and (B) high‑powered (magnification, x100) microscopic images. The section was incubated with ab309139 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) was used. 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-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CUX1+CUX2 with ab309139 at 1/500 (4.78 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat cerebrum with intense staining on superficial cortical layers. (A) Low‑powered (magnification, x40) and (B) high‑powered (magnification, x100) microscopic images. The section was incubated with ab309139 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) was used. 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-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling CUX1+CUX2 with ab309139 at 1/8000 (0.06 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on HEK-293T transfected with a His-tagged mouse CUX2 construct (image A). Positive staining on HEK-293T transfected with a His-tagged mouse CUX1 construct (image B). No staining on HEK-293T transfected with empty plasmid (image C). The section was incubated with ab309139 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) was used. 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-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized mouse primary neuron cells labeling CUX1+CUX2 with ab309139 at 1/50 dilution (1 ug)/Red (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 1/5000 dilution was used as the secondary antibody.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cortex (fresh) tissue labeling CUX1+CUX2 with ab309139 at 1/50 (9.56 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Panel A : merged staining of anti-CUX1+CUX2 (ab309139, green) and anti-NeuN (ab190565, red) on mouse cortex. Panel B : anti-CUX1+CUX2 stained on the mouse cortex. Panel C : anti-NeuN stained in neurons of mouse cortex. The section was incubated in two rounds of staining : in the order of ab309139 and ab190565 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cortex (fresh) tissue labeling CUX1+CUX2 with ab309139 at 1/50 (9.56 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Panel A : merged staining of anti-CUX1+CUX2 (ab309139, green) and anti-NeuN (ab190565, red) on rat cortex. Panel B : anti-CUX1+CUX2 stained on the rat cortex. Panel C : anti-NeuN stained in neurons of rat cortex. The section was incubated in two rounds of staining : in the order of ab309139 and ab190565 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• WB

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Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

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

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The bands beneath the target band (200 kDa) are expected to be degradation products.

In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution.

All lanes:

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (ab309139) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) 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: 200 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution.

All lanes:

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (ab309139) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containi a His-tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human CUX1 expression vector containi a His-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human CUX2 expression vector containi a 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

Observed band size: 200 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Exposure time : Lane 1 : 10 seconds; Lane 2-3 : 40 seconds

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

All lanes:

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (ab309139) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 3:

Rat cerebellum tissue 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 1/20000 dilution

Observed band size: 200 kDa

false

• WB

Supplier Data

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : small intestine tissue(PMID : 8879483). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution.

All lanes:

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (ab309139) at 1/1000 dilution

Lane 1:

Mouse cerebral cortex tissue lysate at 20 µg

Lane 2:

Mouse small intestine tissue lysate at 20 µg

Lane 3:

Rat cerebral cortex tissue 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: 200 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (AB309139)

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

Low expression : Liver tissue(PMID : 8879483).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution.

All lanes:

Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (ab309139) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse testis tissue 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: 200 kDa

true

Exposure time: 180s