Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)

Rabbit Recombinant Monoclonal BRN3A antibody. Carrier free. Suitable for WB, IHC-P, IP, ICC/IF and reacts with Transfected cell lysate - Human, Mouse, Rat samples.

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Images

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	IP	Flow Cyt (Intra)	ICC/IF	IHC-Fr
Human	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Mouse	Tested	Tested	Expected	Not recommended	Tested	Not recommended
Rat	Tested	Tested	Tested	Not recommended	Tested	Not recommended
Transfected cell lysate - Human	Tested	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell lysate - Human, Mouse, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Transfected cell lysate - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell lysate - Human, Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat, Transfected cell lysate - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Transfected cell lysate - Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Transfected cell lysate - Human, Human	Dilution info -	Notes -

Target data

See full target information POU4F1

Function

Multifunctional transcription factor with different regions mediating its different effects. Acts by binding (via its C-terminal domain) to sequences related to the consensus octamer motif 5'-ATGCAAAT-3' in the regulatory regions of its target genes. Regulates the expression of specific genes involved in differentiation and survival within a subset of neuronal lineages. It has been shown that activation of some of these genes requires its N-terminal domain, maybe through a neuronal-specific cofactor. Activates BCL2 expression and protects neuronal cells from apoptosis (via the N-terminal domain). Induces neuronal process outgrowth and the coordinate expression of genes encoding synaptic proteins. Exerts its major developmental effects in somatosensory neurons and in brainstem nuclei involved in motor control. Stimulates the binding affinity of the nuclear estrogene receptor ESR1 to DNA estrogen response element (ERE), and hence modulates ESR1-induced transcriptional activity. May positively regulate POU4F2 and POU4F3. Regulates dorsal root ganglion sensory neuron specification and axonal projection into the spinal cord. Plays a role in TNFSF11-mediated terminal osteoclast differentiation. Negatively regulates its own expression interacting directly with a highly conserved autoregulatory domain surrounding the transcription initiation site. Isoform 2. Able to act as transcription factor, cannot regulate the expression of the same subset of genes than isoform 1. Does not have antiapoptotic effect on neuronal cells.

Additional Targets

POU4F2, POU4F3

Alternative names

BRN3A, RDC1, POU4F1, Brain-specific homeobox/POU domain protein 3A, Homeobox/POU domain protein RDC-1, Oct-T1, Brain-3A, Brn-3A

Recommended products

Rabbit Recombinant Monoclonal BRN3A antibody. Carrier free. Suitable for WB, IHC-P, IP, ICC/IF and reacts with Transfected cell lysate - Human, Mouse, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR26313-54
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

ab317493 is the carrirer-free version of Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

Western blot - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 (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 (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] (Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492) 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 with NFDM/TBST
Lane 2: 293T transfected with a human BRN3A protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with NFDM/TBST
Lane 3: 293T transfected with a human BRN3B protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with NFDM/TBST
Lane 4: 293T transfected with a human BRN3C protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with NFDM/TBST
Lane 5: 293T transfected with a human POU3F2/Brn-2 protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with NFDM/TBST
Lane 6: 293T transfected with a human POU3F3/Brain1 protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with NFDM/TBST
Lane 7: 293T transfected with a human POU3F4/BRN4 protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with NFDM/TBST
Lane 8: 293T transfected with a human POU3F1/Oct6 protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 20-65 kDa, 15 kDa
Exposure time: 3s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat retina tissue labeling BRN3A + BRN3B + BRN3C with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Positive staining on rat retina. The section was incubated with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 (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
Immunocytochemistry/ Immunofluorescence - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/200 dilution (2.575 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 AlexaFluor®488 Goat anti-Rabbit secondary preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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).
Immunocytochemistry/ Immunofluorescence - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/200 dilution (2.575 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 Anti-MAP2 antibody [HM-2] ab11267 anti-MAP2 (mouse mAb) at 1/1000 dilution (1 μg/ml), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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: Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at a 1/200 dilution (2.575 μg/ml), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at a 1/1000 dilution (2 μg/ml).
-ve control 2: Anti-MAP2 antibody [HM-2] ab11267 at a 1/500 dilution (4 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution (2 μg/ml).
Immunocytochemistry/ Immunofluorescence - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/200 dilution (2.575 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 AlexaFluor®488 Goat anti-Rabbit secondary preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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).
Immunocytochemistry/ Immunofluorescence - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/200 dilution (2.575 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 Anti-MAP2 antibody [HM-2] ab11267 anti-MAP2 (mouse mAb) at 1/1000 dilution (1 μg/ml), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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: Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at a 1/200 dilution (2.575 μg/ml), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at a 1/1000 dilution (2 μg/ml).
-ve control 2: Anti-MAP2 antibody [HM-2] ab11267 at a 1/500 dilution (4 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution (2 μg/ml).
Western blot - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 (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] (Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492) at 1/1000 dilution
Lane 1: Rat E15 brain tissue lysate at 20 µg with NFDM/TBST
Lane 2: Rat kidney tissue lysate at 20 µg with NFDM/TBST
Lane 3: Mouse E14.5 brain tissue lysate at 48 µg with NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 45 kDa, 36 kDa
Western blot - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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] (Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492) at 1/1000 dilution
Lane 1: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg with NFDM/TBST
Lane 2: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg with NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 45 kDa
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling BRN3A + BRN3B + BRN3C with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Negative control: no staining on rat cardiac muscle. The section was incubated with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 (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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling BRN3A + BRN3B + BRN3C with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Negative control: no staining on mouse cardiac muscle. The section was incubated with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 (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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse retina tissue labeling BRN3A + BRN3B + BRN3C with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Positive staining on mouse retina. The section was incubated with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 (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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse E14.5 tissue labeling BRN3A + BRN3B + BRN3C with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/1000 (0.535 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Positive staining on mouse E14.5. The section was incubated with Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 (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
Immunoprecipitation - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] - BSA and Azide free (ab317493)
This data was developed using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] 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 Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Rat E15 brain tissue lysate

Lane 2: Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 IP in Rat E15 brain tissue lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492 in rat E15 brain tissue lysate
All lanes: Immunoprecipitation - Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] (Anti-BRN3A + BRN3B + BRN3C antibody [EPR26313-54] ab317492) at 1/30 dilution
All lanes: Rat E15 brain tissue lysate with NFDM/TBST
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s