All lanes : Anti-RECQ1 antibody (ab22830) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 ugLane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 ug Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 ug with Human RECQ1 peptide (ab25318) at 1 µg/mlLane 4 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 ug with Human RECQ1 peptide (ab25318) at 1 µg/mlSecondaryGoat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilutionPerformed under reducing conditions.
ICC/IF image of ab22830 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22830, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red). Panel A shows localisation of ab22830 to the nuclei, Panel B has the Alexa Fluor® 488 channel removed for comparison.
IHC image of ab22830 staining RECQ1 in Human kidney 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 ab22830, 1µg/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.