Anti-Fos B antibody [EPR23489-90]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR23489-90] (AB252237)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Fos B with ab252237 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive nuclear staining in cancer cells of human breast cancer is observed (PMID : 12602926). The section was incubated with ab252237 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR23489-90] (AB252237)

Immunohistochemical analysis of paraffin-embedded human hippocampus tissue labeling Fos B with ab252237 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive nuclear staining in neurons of human hippocampus is observed (PMID : 29805976). The section was incubated with ab252237 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Fos B antibody [EPR23489-90] (AB252237)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Fos B with ab252237 at 1/500 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cells treated with starvation 16 hours, then 12-O-Tetradecanoylphorbol-13-acetate (200nM) for 4 hours. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Fos B antibody [EPR23489-90] (AB252237)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) was serum starved for 16 hours, then treated with 200nM 12-O-Tetradecanoylphorbol-13-acetate (TPA) for 4 h (Red) / Untreated control (Green) cells labeling Fos B with ab252237 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-Fos B antibody [EPR23489-90] (AB252237)

Fos B was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) serum starved overnight, then treated with 200 nM TPA for 4 hours, whole cell lysate 10 μg with ab252237 at 1/30 dilution (2μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab252237 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa serum starved overnight, then treated with 200 nM TPA for 4 hours, whole cell lysate 10 μg.

Lane 2 : ab252237 IP in HeLa serum starved overnight, then treated with 200 nM TPA for 4 hours, whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab252237 in HeLa serum starved overnight, then treated with 200 nM TPA for 4 hours, whole cell lysate.

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

Exposure time : 5 seconds.

The band around 37kDa is caused by splicing isoform.

All lanes:

Immunoprecipitation - Anti-Fos B antibody [EPR23489-90] (ab252237)

Predicted band size: 36 kDa,87 kDa

false

• WB

Lab

Western blot - Anti-Fos B antibody [EPR23489-90] (AB252237)

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

The band around 37kDa is caused by splicing isoform.

The expression profile observed is consistent with what has been described in the literature (PMID : 9204924).

Exposure time : 37 seconds.

All lanes:

Western blot - Anti-Fos B antibody [EPR23489-90] (ab252237) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) serum starved overnight, whole cell lysate at 20 µg

Lane 2:

HeLa serum starved overnight, then treated with 200 nM 12-o-tetradecanoyl phorbol 13-acetate (TPA, <a href='/en-us/products/biochemicals/phorbol-12-myristate-13-acetate-pma-pkc-activator-ab120297'>ab120297</a>) for 4 hours, 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: 36 kDa

Observed band size: 37 kDa,48 kDa

false