![All lanes : Anti-Brk antibody [2H12B8] (ab52273) at 1/2000 dilutionLane 1 : Cell lysates prepared from Hela cellsLane 2 : Cell lysates prepared from A549 cellsLane 3 : Cell lysates prepared from MCF-7 cellsLysates/proteins at 100 µg per lane.SecondaryHRP-conjugated Goat polyclonal to mouse IgG](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/15673_Brk-Primary-antibodies-ab52273-1.jpg)
All lanes : Anti-Brk antibody [2H12B8] (ab52273) at 1/2000 dilutionLane 1 : Cell lysates prepared from Hela cellsLane 2 : Cell lysates prepared from A549 cellsLane 3 : Cell lysates prepared from MCF-7 cellsLysates/proteins at 100 µg per lane.SecondaryHRP-conjugated Goat polyclonal to mouse IgG
IHC image of ab52273 staining in human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab52273, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
![Overlay histogram showing Caco-2 cells stained with ab52273 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52273, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/15675_Brk-Primary-antibodies-ab52273-3.jpg)
Overlay histogram showing Caco-2 cells stained with ab52273 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52273, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.