Anti-B MyB (phospho T487) 抗体 [EPR2204Y] (ab76009)

Immunohistochemistry analysis of paraffin-embedded Human colon cancer tissue sections labelling B MyB (phospho T487) with ab76009 at 1/8000 dilution. The section was incubated with ab76009 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Nuclear staining on Human colon cancer tissue without alkaline phosphatase treatment (Image A); No signal was detected when tissues were treated with alkaline phosphatase (Image B). The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells treated with Alkaline Phosphatase overnight (Left) and untreated HeLa cells (Right) labeling B MyB (phospho T487) with ab76009 at 1/1000 dilution (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150081) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Blocking buffer and concentration: 5% NFDM/TBST

Diluting buffer and concentration: 5% NFDM/TBST

Immunohistochemistry analysis of paraffin-embedded Human lung cancer tissue sections labelling B MyB (phospho T487) with ab76009 at 1/8000 dilution. The section was incubated with ab76009 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Nuclear staining on Human lung cancer tissue without alkaline phosphatase treatment (Image A); No signal was detected when tissues were treated with alkaline phosphatase (Image B). The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling B MyB (phospho T487) with primary antibody anti-B MyB (phospho T487) (ab76009) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing nuclear staining in HeLa cells and no staining in HeLa cells with Alkaline Phosphatase treatment 37℃ for 1 hour. Anti-alpha Tubulin antibody (DM1A) - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counter stain is DAPI (blue).

Blocking buffer and concentration: 5% NFDM/TBST

Diluting buffer and concentration: 5% NFDM/TBST

Blocking buffer and concentration: 5% NFDM/TBST

Diluting buffer and concentration: 5% NFDM/TBST

Dot blot analysis of B MyB (pT487) phospho peptide (lane 1) and B MyB non-phospho peptide (lane 2) with ab76009 at a 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000.
Blocking and dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes