All lanes : Anti-SAP18 antibody (ab31748) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate with Human SAP18 peptide (ab30465) at 1 µg/mlLysates/proteins at 20 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilutionPerformed under reducing conditions.
ICC/IF image of ab31748 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab31748, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Image courtesy of Human Protein Atlasab31748 stainning SAP18 in paraffin-embedded human cervix, showing a distinct nuclear staining of the surface epithelial (squamous) cells. Tissue sections (4µm) were incubated with ab31748 (1/650 dilution) for 30 minutes at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.ab31748 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org