Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat retina tissue labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on rat retina. The section was incubated with ab317492 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cells labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/200 dilution (2.575 μg/ml), followed by ab150081 AlexaFluor®488 Goat anti-Rabbit secondary preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Confocal image showing nuclear staining in mouse primary neurons (shown in green). Counterstained with ab11267 anti-MAP2 (mouse mAb) at 1/1000 dilution (1 μg/ml), followed by ab150120 AlexaFluor®594 Goat anti-Mouse secondary at 1/1000 dilution (2 μg/ml) (Magenta). Nuclear DNA was labelled with DAPI (shown in blue). Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -ve control 1 : ab317492 at a 1/200 dilution (2.575 μg/ml), followed by ab150120 at a 1/1000 dilution (2 μg/ml). -ve control 2 : ab11267 at a 1/500 dilution (4 μg/ml), followed by ab150081 at a 1/1000 dilution (2 μg/ml).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat primary neural/glia cells labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/200 dilution (2.575 μg/ml), followed by ab150081 AlexaFluor®488 Goat anti-Rabbit secondary preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Confocal image showing nuclear staining in rat primary neurons (shown in green). Counterstained with ab11267 anti-MAP2 (mouse mAb) at 1/1000 dilution (1 μg/ml), followed by ab150120 AlexaFluor®594 Goat anti-Mouse secondary at 1/1000 dilution (2 μg/ml) (Magenta). Nuclear DNA was labelled with DAPI (shown in blue). Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -ve control 1 : ab317492 at a 1/200 dilution (2.575 μg/ml), followed by ab150120 at a 1/1000 dilution (2 μg/ml). -ve control 2 : ab11267 at a 1/500 dilution (4 μg/ml), followed by ab150081 at a 1/1000 dilution (2 μg/ml).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse retina tissue labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse retina. The section was incubated with ab317492 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse E14.5 tissue labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse E14.5. The section was incubated with ab317492 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Negative control : no staining on mouse cardiac muscle. The section was incubated with ab317492 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Negative control : no staining on rat cardiac muscle. The section was incubated with ab317492 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse splenocytes labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/200 dilution (2.575 μg/ml), followed by ab150081 AlexaFluor®488 Goat anti-Rabbit secondary preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (2.5 μg/ml) (magenta). Confocal image showing negative staining in mouse splenocytes. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Negative control：splenocyte (PMID : 11470235).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat splenocytes labeling BRN3A + BRN3B + BRN3C with ab317492 at 1/200 dilution (2.575 μg/ml), followed by ab150081 AlexaFluor®488 Goat anti-Rabbit secondary preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (2.5 μg/ml) (magenta). Confocal image showing negative staining in rat splenocytes. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Negative control：splenocyte (PMID : 11470235).

• IP

Supplier Data

Immunoprecipitation - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

BRN3A + BRN3B + BRN3C was immunoprecipitated from 0.35 mg Rat E15 brain tissue lysate with ab317492 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317492 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat E15 brain tissue lysate
Lane 2 : ab317492 IP in Rat E15 brain tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317492 in rat E15 brain tissue lysate

All lanes:

Immunoprecipitation - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] (<a href='/en-us/products/primary-antibodies/brn3a-brn3b-brn3c-antibody-epr26313-54-ab317492'>ab317492</a>) at 1/30 dilution

All lanes:

Rat E15 brain tissue 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: 180s

• WB

Supplier Data

Western blot - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

The lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

The identity of the bands lower than 30 kDa are unknown

All lanes:

Western blot - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] (<a href='/en-us/products/primary-antibodies/brn3a-brn3b-brn3c-antibody-epr26313-54-ab317492'>ab317492</a>) 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

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: 45 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

Negative control : kidney (PMID : 1357630, 11470235)

The identity of the bands lower than 30 kDa in lane 2 and lane 3 are unknown.

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

Exposure time : Lanes 1-2 : 180 seconds; Lane 3 : 92 seconds.

All lanes:

Western blot - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] (<a href='/en-us/products/primary-antibodies/brn3a-brn3b-brn3c-antibody-epr26313-54-ab317492'>ab317492</a>) at 1/1000 dilution

Lane 1:

Rat E15 brain tissue lysate at 20 µg

Lane 2:

Rat kidney tissue lysate at 20 µg

Lane 3:

Mouse E14.5 brain tissue lysate at 48 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: 45 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (AB317493)

This data was developed using ab317492, the same antibody clone in a different buffer formulation.

This antibody recognizes BRN3A, BRN3B and BRN3C.

This antibody does not recognize POU3F2/Brn-2, POU3F3/Brain1, POU3F4/BRN4 and POU3F1/Oct6.

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-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] (<a href='/en-us/products/primary-antibodies/brn3a-brn3b-brn3c-antibody-epr26313-54-ab317492'>ab317492</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 10 µg

Lane 2:

293T transfected with a human BRN3A protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 3:

293T transfected with a human BRN3B protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 4:

293T transfected with a human BRN3C protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 5:

293T transfected with a human POU3F2/Brn-2 protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 6:

293T transfected with a human POU3F3/Brain1 protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 7:

293T transfected with a human POU3F4/BRN4 protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 8:

293T transfected with a human POU3F1/Oct6 protein expression vector containing a myc-His-tag®, whole cell lysate at 10 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: 20-65 kDa,15 kDa

false

Exposure time: 3s