Anti-BRN3A 抗体 [EPR23257-285] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-BRN3A antibody [EPR23257-285] - BSA and Azide free (AB273099)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol-permeabilized Daudi (Human Burkitt's lymphoma lymphoblast (Left) / MOLT-4 (Human lymphoblastic leukemia T lymphoblast (Right) cells labelling BRN3A with ab245230 at 1/400 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : Daudi (PMID : 20460523).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245230).

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-BRN3A antibody [EPR23257-285] - BSA and Azide free (AB273099)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse E14.5 embryo tissue labeling BRN3A with ab245230 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Nuclear staining on mouse E14.5 embryonic brain is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245230).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-BRN3A antibody [EPR23257-285] - BSA and Azide free (AB273099)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse retina tissue labeling BRN3A with ab245230 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Nuclear staining on ganglion cell layer of mouse retina is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245230).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A antibody [EPR23257-285] - BSA and Azide free (AB273099)

Immunohistochemical analysis of paraffin-embedded rat E14.5 tissue labeling BRN3A with ab245230 at 1/2000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on dorsal root ganglia of rat E14.5 (PMID : 8341591). The section was incubated with ab245230 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245230).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRN3A antibody [EPR23257-285] - BSA and Azide free (AB273099)

Immunohistochemical analysis of paraffin-embedded mouse E14.5 tissue labeling BRN3A with ab245230 at 1/2000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on cochleovestibular ganglion and colliculus of mouse E14.5 (PMID : 24709358). The section was incubated with ab245230 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245230).

• IP

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Immunoprecipitation - Anti-BRN3A antibody [EPR23257-285] - BSA and Azide free (AB273099)

BRN3A was immunoprecipitated from 0.35 mg mouse E14.5 brain tissue lysate with ab245230 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab245230 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse E14.5 brain tissue lysate 10μg.

Lane 2 : ab245230 IP in mouse E14.5 brain tissue lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab245230 in mouse E14.5 brain tissue lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

This blot was developed using a higher sensitivity ECL substrate.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245230).

All lanes:

Immunoprecipitation - Anti-BRN3A antibody [EPR23257-285] (<a href='/products/primary-antibodies/brn3a-antibody-epr23257-285-ab245230'>ab245230</a>)

Predicted band size: 43 kDa

Observed band size: 42 kDa

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