IHC image of Rad50 staining in Human colon adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab87918, 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.
![All lanes : Anti-Rad50 antibody [20B5] (ab87918) at 5 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : HT 1080 (Human fibrosarcoma) Whole Cell Lysate Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 5 : HeLa Whole Cell Lysate - Irradiated (5Gy) Lane 6 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate Lane 7 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate Lane 8 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 9 : STO (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 10 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate Lane 11 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate Lysates/proteins at 20 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed und](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/20033_Rad50-Primary-antibodies-ab87918-3.jpg)
All lanes : Anti-Rad50 antibody [20B5] (ab87918) at 5 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : HT 1080 (Human fibrosarcoma) Whole Cell Lysate Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 5 : HeLa Whole Cell Lysate - Irradiated (5Gy) Lane 6 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate Lane 7 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate Lane 8 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 9 : STO (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 10 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate Lane 11 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate Lysates/proteins at 20 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed und
ab87918 (1/200) staining RAD50 in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please see Abreview.See Abreview
![Overlay histogram showing HT1080 cells stained with ab87918 (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 (ab87918, 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 IgG1 [ICIGG1] (ab91353, 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_4/20035_Rad50-Primary-antibodies-ab87918-9.jpg)
Overlay histogram showing HT1080 cells stained with ab87918 (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 (ab87918, 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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.